Consumption and inhibition of plasma cell-like dendritic cells

Abstract

Disclosed are antibodies or fragments thereof, compositions, and methods of use that relate to depleting, inhibiting, or depleting and inhibiting plasma-like dendritic cells (pDCs).

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 395,003, filed on August 4, 2022, the contents of which are incorporated herein by reference in their entirety.

[0003] Statement regarding sequence listing

[0004] This application incorporates by reference the materials in the ST.26 XML file named NPBUS-2_ST.26.xml, which was created on August 4, 2022 and is 20KB in size. Technical Field

[0005] The present disclosure provides novel antibodies and compositions and uses thereof. Background Art

[0006] Plasmacytoid dendritic cells (pDCs) are associated with a variety of disorders. US Patent No. 9,670,283 discloses compositions that can help deplete pDCs in patients in need thereof. Summary of the invention

[0007] The present disclosure provides antibodies or fragments thereof, which comprise a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0008] In an embodiment, the antibody or fragment thereof binds to the target protein with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binds to the epitope having SEQ ID NO:11.

[0009] In an embodiment, the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0010] In an embodiment, the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0011] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises an amino acid sequence selected from SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0012] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises an amino acid sequence selected from SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0013] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises an amino acid sequence selected from SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0014] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises an amino acid sequence selected from SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0015] In an embodiment, the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 5, or a sequence at least about 90% identical to SEQ ID NO: 5.

[0016] In an embodiment, the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 6, or a sequence at least about 90% identical to SEQ ID NO: 6.

[0017] In an embodiment, the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 7, or a sequence at least about 90% identical to SEQ ID NO: 7.

[0018] In an embodiment, the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 8, or a sequence at least about 90% identical to SEQ ID NO: 8.

[0019] In an embodiment, the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 9, or a sequence at least about 90% identical to SEQ ID NO: 9.

[0020] In an embodiment, the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10.

[0021] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 5, or a sequence at least about 90% identical to SEQ ID NO: 5.

[0022] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0023] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0024] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0025] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 6, or a sequence at least about 90% identical to SEQ ID NO: 6.

[0026] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 7, or a sequence at least about 90% identical to SEQ ID NO: 7.

[0027] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 8, or a sequence at least about 90% identical to SEQ ID NO: 8.

[0028] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 9, or a sequence at least about 90% identical to SEQ ID NO: 9.

[0029] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10.

[0030] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0031] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0032] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0033] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0034] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0035] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0036] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0037] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0038] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0039] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0040] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0041] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0042] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0043] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0044] In an embodiment, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0045] In embodiments, a method for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient in need thereof comprises administering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDCs, thereby depleting, inhibiting, or depleting and inhibiting pDCs. The antibody or fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0046] In embodiments of the methods for depleting, inhibiting, or depleting and inhibiting pDC, the epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof binds to pDC with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binding epitope.

[0047] In embodiments of the methods of depleting, inhibiting, or depleting and inhibiting pDC, the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0048] In embodiments of the methods of depleting, inhibiting, or depleting and inhibiting pDC, the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0049] In embodiments of the methods of depleting, inhibiting, or depleting and inhibiting pDCs, the BDCA2 is human BDCA2.

[0050] In embodiments, a method of treating a disorder associated with plasmacytoid dendritic cells (pDCs) in a patient in need thereof comprises administering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen 2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDCs, thereby treating the disorder. The antibody or fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0051] In embodiments of the method of treating a disorder associated with pDCs, the disorder is selected from an infectious disease, an autoimmune disease, an inflammatory disorder, a respiratory disorder, a skin disorder, cancer, and a combination of two or more thereof.

[0052] In an embodiment of the method of treating a disorder associated with pDCs, the disorder is an infectious disease associated with infection with an infectious agent.

[0053] In embodiments of the method of treating a pDC-associated disorder, the infectious agent is one or more viruses or one or more microorganisms.

[0054] In an embodiment of the method of treating a disorder associated with pDC, the infectious agent is a virus selected from the group consisting of: Hepadnaviridae; Flaviviridae; Retroviridae; Herpesviridae; Papovaviridae; Rhabdoviridae; Paramyxoviridae; Reoviridae; Bunyaviridae; Filoviridae; Adenoviridae; Parvoviridae; Arenaviridae; Orthomyxoviridae; Poxviridae; Togaviridae; Coronaviridae; Picornaviridae; Rhinovirus; Orbivirus; Picodnavirus; Encephalomyocarditis virus (EMV); Parainfluenza virus; Adenovirus; Coxsackievirus; Echovirus; Measles virus; Rubella virus; Human papillomavirus; Canine distemper virus; Canine infectious hepatitis virus; Feline calicivirus; Feline rhinotracheitis virus; TGE virus (swine); Foot-and-mouth disease virus; Simian virus 5; Human parainfluenza virus type 2; Human subpneumovirus; and Enterovirus.

[0055] In an embodiment of the method of treating a disorder associated with pDC, the infectious agent is a microorganism selected from the group consisting of Rickettsia, Chlamydia, Mycobacteria, Clostridia, Corynebacteria, Mycoplasma, Ureaplasma, Legionella, Shigella, Salmonella, pathogenic Escherichia coli species, Bordatella, Neisseria, Treponema, Bacillus, Haemophilus, Moraxella, Vibrio, Staphylococcus spp., Streptococcus spp., Campylobacter spp. spp.), Borrelia spp., Leptospira spp., Erlichia spp., Klebsiella spp., Pseudomonas spp., and Helicobacter pylori spp.

[0056] In an embodiment of the method of treating a disorder associated with pDC, the infectious agent is a microorganism selected from the group consisting of Helicobacter pylori, Chlamydia pneumoniae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Treponema pallidum. Bacillus anthracis, Salmonella typhi, Vibrio cholera. pestis (Pasteurella pestis), (Yersinia pestis), Pseudomonas aeruginosa, Campylobacter jejuni, Clostridium dificile, Clostridium botulinum, Mycobacterium tuberculosis, Borrelia burgdorferi, Haemophilus ducreyi, Corynebacterium diphtheria, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenza, and enterotoxic Escherichia coli.

[0057] In an embodiment of the method of treating a disorder associated with pDCs, the disorder comprises an autoimmune disease selected from the group consisting of diabetes, cutaneous psoriasis, hyperthyroidism, autoimmune adrenal insufficiency, hemolytic anemia, rheumatic myocarditis, systemic lupus erythematosus, cutaneous lupus erythematosus, psoriatic arthritis, Sjögren's syndrome polymyositis, myositis, multiple sclerosis, Crohn's disease, ulcerative colitis, lupus, inflammatory bowel syndrome, dermatomyositis, dermatitis, atopic dermatitis, scleroderma, polymyositis, psoriasis, alopecia areata, rheumatoid arthritis, graft versus host disease, interferon disease, irritable bowel syndrome, and a combination of two or more of these.

[0058] In an embodiment of the method of treating a disorder associated with pDCs, the disorder comprises an inflammatory disorder comprising acute kidney injury.

[0059] In embodiments of the method of treating a disorder associated with pDCs, the disorder comprises a respiratory disorder comprising asthma, pulmonary fibrosis, or a combination of asthma and pulmonary fibrosis.

[0060] In embodiments of the method of treating a disorder associated with pDCs, the disorder comprises a skin disorder selected from scleroderma, psoriasis, atopic dermatitis, dermatomyositis, and a combination of two or more thereof.

[0061] In an embodiment of the method of treating a disorder associated with pDC, the disorder comprises a cancer selected from the group consisting of breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head and neck cancer, breast cancer, ovarian cancer, lung cancer, small cell lung cancer, Wilms tumor, cervical cancer, testicular cancer, bladder cancer, pancreatic cancer, gastric cancer, colon cancer, prostate cancer, genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, B-cell acute The invention further comprises the following: myeloid leukemia, blastic plasmacytoid dendritic cell neoplasm ("BPDCN" or "pDC leukemia"), chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia ("AML"), chronic myeloid leukemia, acute myeloid leukemia, senescence-associated hairy cell leukemia, chronic myelomonocytic leukemia ("CMML"), myelodysplastic syndrome, chronic myelofibrosis, multiple myeloma, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL) (L265P), polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, retinoblastoma, and combinations of two or more thereof.

[0062] In an embodiment of the method of treating a disorder associated with pDC, the epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof binds to the pDC with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binding epitope.

[0063] In embodiments of the method of treating a disorder associated with pDCs, the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0064] In embodiments of the method of treating a disorder associated with pDCs, the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0065] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0066] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0067] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0068] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0069] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 5, or a sequence at least about 90% identical to SEQ ID NO: 5.

[0070] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 6, or a sequence at least about 90% identical to SEQ ID NO: 6.

[0071] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 7, or a sequence at least about 90% identical to SEQ ID NO: 7.

[0072] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 8, or a sequence at least about 90% identical to SEQ ID NO: 8.

[0073] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 9, or a sequence at least about 90% identical to SEQ ID NO: 9.

[0074] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10.

[0075] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0076] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0077] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0078] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0079] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0080] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0081] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:1, or a sequence at least about 90% identical to SEQ ID NO:1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0082] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 9, or a sequence at least about 90% identical to SEQ ID NO: 9.

[0083] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1, or a sequence at least about 90% identical to SEQ ID NO: 1; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO: 10, or a sequence at least about 90% identical to SEQ ID NO: 10.

[0084] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0085] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0086] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0087] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0088] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0089] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0090] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0091] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0092] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0093] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0094] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0095] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0096] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:2, or a sequence at least about 90% identical to SEQ ID NO:2; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0097] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:3, or a sequence at least about 90% identical to SEQ ID NO:3; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0098] In an embodiment of the method of treating a disorder associated with pDC, the heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and the light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0099] In an embodiment, a kit for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient in need thereof comprises: instructions; and an antibody or fragment thereof. The antibody or fragment thereof comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; the light chain variable region comprising an amino acid sequence selected from SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0100] In an embodiment, the pharmaceutical composition comprises: at least one excipient; and an antibody or fragment thereof. The antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0101] In embodiments, the antibodies or fragments thereof may be used therapeutically.

[0102] In embodiments, the antibodies or fragments thereof can be used to deplete, inhibit, or deplete and inhibit plasmacytoid dendritic cells (pDCs).

[0103] In an embodiment, the antibody or fragment thereof may be used in the preparation of a medicament.

[0104] The present disclosure is not intended to identify the key or essential features of the claimed subject matter, nor is it intended to be used as an auxiliary means to limit the scope of the claimed subject matter. Further embodiments, forms, features and aspects of the present disclosure will become apparent from the description and drawings provided herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0105] The teachings of some embodiments of the present disclosure will be better understood by reference to the description taken in conjunction with the accompanying drawings, in which:

[0106] Figure 1 shows a schematic diagram of expression vectors for producing chimeric and humanized anti-BDCA2 antibodies according to an embodiment;

[0107] Figure 2 shows SDS-PAGE analysis of NPB-303 according to an embodiment;

[0108] FIG3 shows protein analysis of NPB-303 on a Superose 6 column using an AKTA FPLC system according to an embodiment; Figure 3A , BioRad gel filtration standard; B.NPB-303.

[0109] Figure 4 Biophysical binding of 15b, 12b, and NPB-303 to BDCA2 antigen from monkey (panel A) or human (panel B), according to the embodiments, is shown;

[0110] Figure 5 A structural model of the BDCA2 extracellular domain according to an embodiment is shown; green: BDCA2 extracellular domain; yellow: NPB-303 epitope; orange: BDCA2 lectin (ligand) binding residues.

[0111] Figure 6 A structural model of the variable (binding) domain of NPB-303 according to an embodiment is shown; green: VH framework region; yellow: VH CDR; blue: VL framework region; brown: VL CDR; red: aspartate isomerization motif.

[0112] Figure 7 shows binding characterization of NPB-303 antibody by surface plasmon resonance (SPR) according to an embodiment;

[0113] Figure 8 shows the in vitro binding characterization of NPB-303 mAb to human BDCA2 antigen according to the embodiments;

[0114] Fig. 9 shows characterization of binding of NPB-303 mAb to human pDC in vitro according to an embodiment;

[0115] Fig.10 shows a FACS graph in which NPB-303 inhibits pDC IFNα response to CpG-A stimulation in vitro according to an embodiment;

[0116] Fig.11 shows a dose response curve of NPB-303 relative to IFNα according to an embodiment;

[0117] Fig.12 The IC values ​​of NPB-303 in multiple human donor PBMC samples are shown according to the embodiments. 50 ;

[0118] Fig.13 shows an in vitro efficacy analysis of NPB-303 according to an embodiment;

[0119] Fig.14 shows the pharmacokinetic profile of NPB-303 in genetically modified mice according to the embodiments;

[0120] Fig.15shows a FACS graph demonstrating NPB-303-induced pDC depletion and BDCA2 downregulation in blood according to an embodiment;

[0121] Fig.16 Summary data showing pDC depletion (% CD4+CD123+ of Lin-) and BDCA2 downregulation on pDC (% CD4+BDCA2+ of Lin-) are shown according to an embodiment;

[0122] Fig.17 shows the in vivo pharmacodynamic profile of NPB-303 mAb in humanized mice according to the embodiments;

[0123] Fig.18 A summary of the percentage of pDCs or percentage of BDCA2+ cells in humanized mice treated with an isotype control or NPB-303 (normalized to the isotype control antibody) according to the embodiments is shown;

[0124] Fig.19 Shown is a summary analysis of human BDCA2+ pDC in humanized mouse spleen cells according to the embodiments;

[0125] Fig. 20 Shown is a summary analysis of human CD123+pDC in humanized mouse spleen cells according to the embodiment;

[0126] Fig.21 A summary of other cell types (Lin-CD4-, CD4+CD123-, and CD4-CD123-) in the blood of humanized mice treated with isotype control mAb or NPB-303 according to the embodiments is shown;

[0127] Fig. 22 Endotoxin testing of NPB-303 (1:1000 dilution) using a kinetic turbidimetric and kinetic chromogenic test (ENDOSAFE) is shown;

[0128] Fig.23 According to the embodiment, NPB-303 or Summary of antitumor activity in mice bearing human immune and tumor cells (“HIT mice”);

[0129] Fig.24 shows a summary of the body weights of tumor-bearing mice in each treatment group according to the embodiment;

[0130] Fig.25 According to an embodiment, NPB-303 but not Summary of pDC depletion in spleen;

[0131] Fig.26 According to an embodiment, NPB-303 but not Summary of pDC depletion leading to tumors;

[0132] Fig. 27 According to an embodiment, NPB-303 but not Summary of the downregulation of BDCA2 in the spleen;

[0133] Fig.28 According to an embodiment, NPB-303 but not Summary of causes of BDCA2 downregulation in tumors;

[0134] Fig.29 A schematic diagram of a method for establishing a tumor-bearing humanized mouse model according to an embodiment is shown;

[0135] Fig.30 shows representative flow cytometry plots of human pDCs in tumors isolated from HIT-TNBC mice according to the embodiments;

[0136] Fig.31 A summary of the percentage of human pDCs in tumors according to the embodiments is shown;

[0137] Fig.32 shows representative flow cytometry plots of tumors isolated from HIT-TNBC mice and human T cells in the blood according to the embodiments;

[0138] Fig.33 shows the mean fluorescence intensity (MFI) of Tim3 and PD1 on CD4+T cells according to the embodiment;

[0139] Fig.34 shows the mean fluorescence intensity (MFI) of Tim3 and PD1 on CD8+ T cells according to the embodiment;

[0140] Fig.35 Showing the dose-dependent anti-tumor activity (measured by tumor volume) of NPB-303 according to the embodiments;

[0141] Fig.36 shows stable body weight over time for HIT mice treated with NPB-303 according to an embodiment;

[0142] Fig.37 Shown are representative flow cytometry plots and summary of the percentage of CD4+CD123+ of Lin- in spleen according to an embodiment;

[0143] Fig.38Shown are representative flow cytometry plots and a summary of the percentage of CD4+CD123+ of Lin- in tumors according to the embodiments;

[0144] Fig.39 shows tumor volume over time for a second human TNBC cell line (MDA-MB-468) according to an embodiment;

[0145] Fig.40 Showing the dose-dependent anti-tumor activity (measured by tumor volume) of NPB-303 according to the embodiments;

[0146] Fig.41 shows stable body weight over time of HIT mice treated with NPB-303 according to an embodiment;

[0147] Fig.42 NPB-303 according to the embodiment is shown with Research procedures for combination therapies;

[0148] Fig.43 NPB-303 alone, antitumor activity (measured by tumor volume) of individual and combination therapies;

[0149] Fig.44 According to the embodiment, NPB-303 alone, Stable body weight over time in HIT mice treated with single and combination therapies;

[0150] Fig.45 It is shown that according to the embodiments, PD1 is downregulated in tumor-infiltrating CD8+ cells in both the NPB-303 group and the combination therapy group compared to the isotype control group;

[0151] Fig.46 It is shown that according to the embodiment, Tim3 is downregulated in tumor-infiltrating CD8+ cells in both the NPB-303 group and the combination therapy group compared to the isotype control group;

[0152] Fig.47 The results from each treatment group (isotype control group, NPB-303 alone, Representative liver images of patients with NPB-303 alone and in combination therapy (Figure 2A), demonstrating that, according to the embodiments, NPB-303 alone or NPB-303 and There was a reduction in tumor nodules in the group of mice treated with the combination;

[0153] Fig.48 Each treatment group (isotype control group, NPB-303 alone, Summary of liver infiltration scores for 100 mg / kg / day, alone and in combination therapy);

[0154] Fig.49 Each treatment group (isotype control group, NPB-303 alone, Representative H&E staining data of liver tissue histological sections (100 μm) of 100 μm and 20 μm of 100 μm treated with 100 μm daptomycin alone and combination therapy;

[0155] Fig.50 Representative lung H&E staining images (3× and 17×) are shown according to an embodiment;

[0156] Fig.51 Each treatment group (isotype control group, NPB-303 alone, Summary of lung infiltration scores for 24-week follow-up (alone and combination therapy);

[0157] Fig.52 The thermal denaturation (T m ) and the occurrence of aggregation (T agg );

[0158] Fig.53 shows the binding activity of NPB-303 under different storage temperature conditions according to the embodiment;

[0159] Fig.54 Detailed sequence modifications of the NPB-303 antibody according to the embodiments are shown;

[0160] Fig.55 shows a sensorgram of SPR-based affinity testing for 24 antibodies according to an embodiment;

[0161] Fig.56 A graph showing a set of IFNα inhibition assays according to an embodiment;

[0162] Fig.57 The efficacy of the antibodies according to the embodiments is shown (IC 50 ) summary;

[0163] Fig.58 shows a sensorgram of SPR-based affinity testing for three antibodies according to an embodiment;

[0164] Fig.59 shows a set of graphs demonstrating dose-dependent inhibition of TLR-9-induced IFNα in PBMCs from two healthy human donors using NPB-303 and three additional monoclonal antibodies, according to an embodiment;

[0165] Fig.60The efficacy of the antibodies according to the embodiments on IFNα inhibition (IC 50 ) summary;

[0166] Fig.61 A set of representative FACS plots of human pDCs (CD4+CD123+) and CD4+BDCA2+ cells (gated on human CD45+ and then Lin- cells) in the blood of humanized mice treated with isotype control antibody, NPB-303, and 3 other antibodies, according to the embodiments;

[0167] Fig.62 shows a summary of the pDC percentage of Lin- cells in the blood of humanized mice treated with isotype control antibody, NPB-303 and 3 other antibodies according to the embodiments;

[0168] Fig.63 shows a summary of the BDCA2+ percentage of Lin- cells in the blood of humanized mice treated with an isotype control antibody, NPB-303, and 3 other antibodies, according to the embodiments;

[0169] Fig.64 A set of representative FACS plots of human pDC (CD4+CD123+) and CD4+BDCA2+ cells (gated on human CD45+ then Lin- cells) in the spleen of humanized mice treated with isotype control antibody, NPB-303 and 3 other antibodies according to the embodiments are shown;

[0170] Fig.65 Shown is a summary of the pDC percentage of Lin- cells in the spleen of humanized mice treated with isotype control antibody, NPB-303 and 3 other antibodies according to the embodiments;

[0171] Fig.66 shows a summary of the BDCA2+ percentage of Lin- cells in the spleen of humanized mice treated with an isotype control antibody, NPB-303, and 3 other antibodies, according to the embodiments;

[0172] Fig.67 A set of representative FACS graphs of myeloid cells (Lin-CD4-), conventional DCs (CD4+CD123-), and other cell types (Lin-CD4-CD123-) in the blood of humanized mice treated with isotype control antibodies, NPB-303, and three other antibodies according to the embodiments are shown;

[0173] Fig.68shows a summary of the percentages of Lin-CD4-, CD4+CD123-, and Lin-CD4-CD123- cells in the blood of humanized mice treated with an isotype control antibody, NPB-303, and three other antibodies, according to the embodiments; and

[0174] Fig.69 Shown is a summary of the percentages of Lin-CD4-, CD4+CD123-, and Lin-CD4-CD123- cells in the spleens of humanized mice treated with an isotype control antibody, NPB-303, and three other antibodies, according to the embodiments. DETAILED DESCRIPTION

[0175] The following embodiments are not intended to be exhaustive or to limit the invention to the precise forms disclosed in the following detailed description. Instead, these embodiments are selected and described to enable others skilled in the art to appreciate and understand the principles and practices of the present disclosure.

[0176] Unless the context indicates otherwise, it is specifically intended that the various features of the invention described herein can be used in any combination. In addition, the present invention also contemplates that in some embodiments of the present invention, any feature or combination of features described herein can be excluded or omitted. For example, if the specification indicates that a composite comprises components A, B, and C, it is specifically intended that any one of A, B, or C, or a combination thereof, can be omitted and excluded, alone or in any combination.

[0177] Amino acids are referred to herein in the manner recommended by the IUPAC-IUB Biochemical Nomenclature Commission, or by either the one-letter code or the three-letter code, both consistent with 37 CFR §1.822 and established usage.

[0178] The present disclosure provides antibodies or fragments thereof, which comprise a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0179] As used in the specification and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. Similarly, as used herein, "and / or" refers to and encompasses any and all possible combinations of one or more of the relevant listed items, as well as the lack of combination when interpreted in an alternative manner (or). As used herein, the term "about" is intended to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount when referring to a measurable value (e.g., the amount of a polypeptide, dosage, time, temperature, enzyme activity or other biological activity, etc.).

[0180] The transitional phrase "consisting essentially of" means that the scope of a claim is interpreted to cover the specific materials or steps described in the claim, "as well as materials or steps that do not materially affect the basic, novel characteristics of the claimed subject matter." See, In re Herz, 537 F.2d 549, 551-52, 190 USPQ 461, 463 (CCPA 1976) (emphasis in the original); see also MPEP §2111.03 (9th Ed., 10th Rev.). The term "consisting essentially of" (and grammatical variations thereof), when applied to a polypeptide sequence, refers to a polypeptide consisting of the sequence (e.g., SEQ ID NO) and a total of ten or fewer (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) additional amino acids at the N-terminus and / or C-terminus of the sequence, such that the function of the polypeptide is not substantially altered. The additional amino acids include the total number of additional amino acids at both ends added together. When applied to polypeptides, the term "substantially altered" means that the epitope binding activity is increased or decreased by at least about 50% or more compared to the activity of a polypeptide consisting of the sequence.

[0181] The term "isolated" may refer to a nucleic acid, nucleotide sequence or polypeptide that is substantially free of cellular material, viral material and / or culture medium (when produced by recombinant DNA techniques) or chemical precursors or other chemicals (when chemically synthesized). In addition, an "isolated fragment" refers to a fragment of a nucleic acid, nucleotide sequence or polypeptide that is not a naturally occurring fragment and is not found in the natural state. "Isolated" does not mean that the preparation is technically pure (homogeneous), but that its purity is sufficient to provide the polypeptide or nucleic acid in a form that can be used for the intended purpose.

[0182] The term "fragment" when applied to a polypeptide may be understood as an amino acid sequence that is shortened in length relative to a reference polypeptide or amino acid sequence, and includes, consists essentially of, and / or consists of an amino acid sequence of contiguous amino acids that are identical or nearly identical (e.g., about 90%, about 92%, about 95%, about 98%, about 99% identical) to a reference polypeptide or amino acid sequence. If appropriate, such a polypeptide fragment according to the present disclosure may be incorporated into a larger polypeptide of which it is a component. In some embodiments, such a fragment may include, consist essentially of, and / or consist of a peptide having a length of at least about 4, 6, 8, 10, 12, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200 or more contiguous amino acids of a polypeptide or amino acid sequence.

[0183] As used herein, the terms "protein" and "polypeptide" are used interchangeably and encompass both peptides and proteins, unless otherwise indicated.

[0184] As used herein, a "functional polypeptide" or "functional fragment" is a polypeptide or fragment that substantially retains at least one biological activity (e.g., target protein binding) that is typically associated with the polypeptide. In a specific embodiment, a "functional polypeptide" or "functional fragment" substantially retains all activities possessed by an unmodified peptide. "Substantially retaining" biological activity means that the polypeptide retains at least about 20%, 30%, 40%, 50%, 60%, 75%, 85%, 90%, 95%, 97%, 98%, 99% or more of the biological activity of the natural polypeptide (and may even have a higher level of activity than the natural polypeptide). A "non-functional polypeptide" is a polypeptide that exhibits little or substantially no detectable biological activity that is typically associated with the polypeptide (e.g., at most only an insignificant amount, e.g., less than about 10% or even less than about 5%). Biological activity (e.g., protein binding) can be measured using assays known to those skilled in the art and described herein.

[0185] Disclosed herein are antibodies that specifically bind to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and deplete, inhibit, or deplete and inhibit pDCs. Such antibodies can be used to deplete, inhibit, or deplete and inhibit pDCs in a patient, for example, for research or therapeutic purposes. Such antibodies can be used to treat conditions associated with pDCs. Thus, one aspect relates to antibodies or fragments thereof that specifically bind to BDCA2 and deplete, inhibit, or deplete and inhibit pDCs when administered to a patient.

[0186] As used herein, the term "antibody" or "antibody" refers to all types of immunoglobulins, including IgG, IgM, IgA, IgD and IgE. Antibodies can be monoclonal or polyclonal and can be from any species, including (for example) mice, rats, rabbits, horses, goats, sheep, camels or humans, or can be chimeric antibodies. For example, see Walker et al., Molec. Immunol. 26: 403 (1989). Antibodies can be recombinant monoclonal antibodies produced according to the methods disclosed in U.S. Patent Nos. 4,474,893 or 4,816,567. Antibodies can also be chemically constructed according to the methods disclosed in U.S. Patent No. 4,676,980.

[0187] Antibody fragments include, for example, Fab, Fab', F(ab')2, and Fv fragments; domain antibodies, diabodies; vaccine antibodies, linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments. Such fragments can be produced by known techniques. For example, F(ab')2 fragments can be produced by pepsin digestion of antibody molecules, while Fab fragments can be produced by reducing the disulfide bonds of F(ab')2 fragments. Alternatively, Fab expression libraries can be constructed to allow rapid and easy identification of monoclonal Fab fragments with desired specificity (Huse et al., Science 254:1275 (1989)).

[0188] Antibodies can be altered or mutated to be compatible with species other than the species in which the antibody was produced. For example, antibodies can be humanized or camelized. When the patient is human, the antibody can be a humanized antibody. For example, see Safdari et al., Antibody humanization methods-a review and update. Biotechnol. Genet. Eng. Rev., 2013; 29: 175-86. The humanized form of a non-human (e.g., mouse) antibody is a chimeric immunoglobulin, an immunoglobulin chain, or a fragment thereof (e.g., Fv, Fab, Fab', F(ab')2, or other antigen-binding subsequences of an antibody) containing minimal sequences from a non-human immunoglobulin. Humanized antibodies include human immunoglobulins (receptor antibodies) in which residues of the complementary determining regions (CDRs) of the receptor are replaced by CDR residues of a non-human species (donor antibody) (e.g., mouse, rat, or rabbit) having the desired specificity, affinity, and ability. In some embodiments, the Fv framework region residues of a human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also include residues that are not present in the receptor antibody or the imported CDR or framework region sequences. In general, humanized antibodies can include at least one, usually substantially all of two variable domains, wherein all or substantially all of the CDR regions correspond to the CDR regions of non-human immunoglobulins, and all or substantially all of the framework (FR) regions (i.e., the sequences between the CDR regions) are framework (FR) regions of human immunoglobulin consensus sequences. Humanized antibodies can also include at least a sub-immunoglobulin constant region (Fc) portion, typically a constant region of a human immunoglobulin (Jones et al., Nature 321:522 (1986); Riechmann et al., Nature, 332:323 (1988); and Presta, Curr. Op. Struct. Biol. 2:593 (1992)).

[0189] As mentioned above, the method of humanizing non-human antibodies is known. In general, humanized antibodies have one or more amino acid residues from non-human sources introduced therein. These non-human amino acid residues are generally referred to as "import" residues, usually taken from "import" variable domains. Humanization can be carried out according to the method of Winter and his colleagues (Jones et al., Nature 321:522 (1986); Riechmann et al., Nature 332:323 (1988); Verhoeyen et al., Science 239:1534 (1988)), by replacing the corresponding sequence of human antibodies with rodent CDR or CDR sequences. Therefore, this "humanized" antibody is a chimeric antibody (U.S. Patent No. 4,816,567), in which the part substantially less than the complete human variable domain is replaced by the corresponding sequence from non-human species. In practice, humanized antibodies may be human antibodies in which some CDR residues (eg, all or part of a CDR) and possibly some FR residues are substituted by residues from analogous sites in rodent (eg, murine) antibodies.

[0190] Human antibodies can be produced using various techniques known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol. 227: 381 (1991); Marks et al., J. Mol. Biol. 222: 581 (1991)). The techniques of Cole et al. and Boerner et al. can also be used to prepare human monoclonal antibodies (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc.: New York, p. 77 (1985) and Boerner et al., J. Immunol. 147: 86 (1991)). Similarly, human antibodies can be prepared by introducing human immunoglobulin loci into transgenic animals (e.g., mice) in which endogenous immunoglobulin genes have been partially or completely inactivated. After challenge, the production of human antibodies is observed, which is very similar to that observed in humans in all aspects, including gene rearrangement, assembly, and antibody repertoires. For example, this method is described in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in the following scientific publications: Marks et al., Bio / Technology 10:779 (1992); Lonberg et al., Nature 368:856 (1994); Morrison, Nature 368:812 (1994); Fishwild et al., Nature Biotechnol. 14:845 (1996); Neuberger, Nature Biotechnol. 14:826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13:65 (1995).

[0191] Polyclonal antibodies for practicing the present disclosure can be produced according to known procedures: immunizing a suitable animal (e.g., rabbit, goat, etc.) with an antigen to which a monoclonal antibody against a target binds, collecting immune serum from the animal, and isolating polyclonal antibodies from the immune serum. The polynucleotide sequence and polypeptide sequence of BDCA2 are known and can be found in sequence databases such as GenBank. Examples of sequences include human BDCA2 polypeptide sequence (Accession No. Q8WTTO) and polynucleotide sequence (Accession No. AF293615), which are incorporated herein by reference in their entirety.

[0192] Monoclonal antibodies can be produced in hybridoma cell lines according to the techniques of Kohler and Milstein Nature 265:495 (1975). For example, a solution containing an appropriate antigen can be injected into mice, and mice can be killed after a sufficient time and spleen cells can be obtained. Splenocytes can then be immortalized by fusing splenocytes with myeloma cells or lymphoma cells (usually in the presence of polyethylene glycol) to produce hybridoma cells. The hybridoma cells are then cultured in a suitable culture medium, and monoclonal antibodies with the desired specificity are screened from the supernatant. Monoclonal Fab fragments can be produced in Escherichia coli by recombinant techniques known to those skilled in the art. For example, see Huse, Science 246:1275 (1989).

[0193] Antibodies specific for the target polypeptide can also be obtained by phage display technology known in the art.

[0194] In some embodiments, the antibodies or fragments thereof formed by the above methods can bind to the target protein with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binds to an epitope having SEQ ID NO: 11. SEQ ID NO: 11 is a fragment of SEQ ID NO: 12, which provides the amino acid sequence of the BDCA2 antigenic epitope. In embodiments, the affinity K d) can be 0.1nM to 90nM, 0.1nM to 80nM, 0.1nM to 70nM, 0.1nM to 60nM, 0.1nM to 50nM, 0.1nM to 40nM, 0.1nM to 30nM, 0.1nM to 20nM, 0.1nM to 10nM, 0.1nM to 9 ...nM, 0.1nM to 7nM, 0.1nM to 6nM, 0.1nM to 0.1nM to 0.5nM, 0.1nM to 0.4nM, 0.1nM to 0.3nM, 0.1nM to 0.2nM, 0.1nM to 0.6nM, 0.1nM to 0.5nM, 0.1nM to 0.4nM, 0.1nM to 0.3nM, 0.1nM to 0.2nM, 0.2nM to 100nM, 0.3nM to 100nM, 0.4nM to 100nM, 0.5nM to 100nM, 0.6nM to 100nM, 0.7nM to 100nM, 0.8nM to 100nM, 0.9nM to 100nM, 1nM to 100nM, 2nM to 100nM, 3nM to 100nM, 4nM to 100nM, 5nM to 100nM In some embodiments, the present invention may be directed to a pharmaceutical composition comprising the following: a) 100 nM to 400 nM, b) 100 nM to 500 nM, c) 100 nM to 500 nM, d) 100 nM to 500 nM, 3) 100 nM to 500 nM, 4) 100 nM to 500 nM, 5) 100 nM to 500 nM, 6) 100 nM to 500 nM, 7) 100 nM to 500 nM, 8) 100 nM to 500 nM, 9) 100 nM to 100 nM, 10) 100 nM to 500 nM, 10) 100 nM to 500 nM, 10) 100 nM to 500 nM,

[0195] The term "affinity" as used herein refers to the degree or strength of binding of an antibody to an epitope. Affinity can be measured and / or expressed in a variety of ways known in the art, including but not limited to the equilibrium dissociation constant (K D or K d ), apparent equilibrium dissociation constant (K D ′ or K d ′), and IC 50 (Amount required to achieve 50% inhibition in a competition assay). It will be understood that affinity is the average affinity of a population of antibodies for a particular epitope. Screening can be performed using various immunoassays to identify antibodies having the desired affinity for the peptide sequence given in SEQ ID NO:11.

[0196] The antibodies can be conjugated to a solid support (e.g., beads, plates, slides, or wells formed from materials such as latex or polystyrene) according to known techniques. The antibodies can also be conjugated to a detectable group, such as a radiolabel (e.g., 35 S. 125 S.131 I), enzyme labels (e.g., horseradish peroxidase, alkaline phosphatase) and fluorescent labels (e.g., fluorescein). In the methods disclosed herein, the formation of antibody / antigen complexes can be determined by detecting, for example, precipitation, agglutination, flocculation, radioactivity, color development or change, fluorescence, luminescence, etc.

[0197] In some embodiments, the CDRs of an antibody may comprise at least one heavy chain variable region and at least one light chain variable region.

[0198] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 1 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0199] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 2 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 2 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0200] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 3 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 3 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0201] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising at least about 50 consecutive amino acids of the amino acid sequence of SEQ ID NO: 4 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0202] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 5 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 5 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0203] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 6 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 6 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0204] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 7 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 7 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0205] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 8 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0206] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 9 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 9 or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0207] In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In some embodiments, the antibody or fragment thereof comprises a light chain variable region comprising at least about 50 consecutive amino acids of an amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical thereto, such as at least about 100, 150 or 200 or more consecutive amino acids.

[0208] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1 or at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0209] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 2 or at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0210] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 3 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0211] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 4 or a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence at least about 90% identical thereto, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0212] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical, to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 5 or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0213] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical, to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 6 or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0214] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical, to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 7 or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0215] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical, to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 8 or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0216] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical, to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 9 or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0217] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical, to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. In addition, the antibody or fragment thereof comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 10 or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0218] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 5, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0219] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:5, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0220] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:5, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0221] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:5, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0222] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0223] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0224] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0225] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0226] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 1, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0227] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 6, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0228] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 7, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0229] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0230] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0231] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0232] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:6, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0233] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:3, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:7, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0234] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 8, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0235] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 9, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0236] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 3, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0237] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:6, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0238] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:7, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0239] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:8, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0240] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:4, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO:9, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0241] In some embodiments, the antibody or fragment thereof comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 4, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto, and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10, or a sequence at least about 90% identical, such as at least about 95%, 96%, 97%, 98% or 99% identical thereto.

[0242] In some embodiments, the above-mentioned antibodies or fragments thereof may be included in a composition. In some embodiments, the composition is a pharmaceutical composition comprising an antibody or fragment thereof in a pharmaceutically acceptable carrier, optionally containing one or more medicaments, pharmaceutical agents, carriers, adjuvants, dispersants, diluents, etc. The term "pharmaceutically acceptable" as used herein refers to a component that is harmless to the patient and is compatible with other ingredients, active ingredients, salts or components with respect to formulation components such as carriers, excipients and diluents. "Pharmaceutically acceptable" includes "veterinarily acceptable", and therefore includes applications in humans and non-human mammals, respectively.

[0243] More specifically, the term "pharmaceutical composition" as used herein refers to a pharmaceutical formulation containing an antibody or fragment thereof of the present disclosure and suitable for administration to a patient for therapeutic purposes. The term "patient" as used herein refers to a living organism treated with an antibody or fragment thereof of the present disclosure, including but not limited to any mammal, such as humans, other primates (e.g., monkeys, chimpanzees, etc.), companion animals (e.g., dogs, cats, horses, etc.), farm animals (e.g., goats, sheep, pigs, cattle, etc.), laboratory animals (e.g., mice, rats, etc.), and wild and zoo animals (e.g., wolves, bears, deer, etc.).

[0244] In some embodiments, the pharmaceutical composition may include at least one pharmaceutically acceptable component to provide an improved formulation of the disclosed compounds, including but not limited to one or more pharmaceutically acceptable carriers, excipients or diluents. The carrier, excipient or diluent may take a variety of forms, depending on the formulation form desired for administration.

[0245] The term "carrier" as used herein includes, but is not limited to, calcium carbonate, calcium phosphate, various sugars (e.g., lactose, glucose or sucrose), various starches, cellulose derivatives, gelatin, lipids, liposomes, nanoparticles, physiologically acceptable liquids as solvents or suspensions (including, for example, sterile solutions of water for injection (WFI), saline solutions, D-glucose solutions, Hank's solutions, Ringer's solutions, vegetable oils, mineral oils, animal oils, polyethylene glycols, liquid paraffin, etc.).

[0246] The term "excipient" as used herein generally includes, but is not limited to, fillers, binders, disintegrants, glidants, lubricants, complexing agents, solubilizers, stabilizers, preservatives and surfactants, which can be selected to help administer the compound by a particular route. Suitable excipients can also include, for example, colloidal silicon dioxide, silica gel, talc, magnesium silicate, calcium silicate, sodium aluminosilicate, magnesium trisilicate, powdered cellulose, microcrystalline cellulose, carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose, sodium benzoate, calcium carbonate, magnesium carbonate, stearic acid, aluminum stearate, calcium stearate, magnesium stearate, zinc stearate, sodium stearyl fumarate, syloid, stearowet C, magnesium oxide, starch, sodium starch glycolate, glyceryl monostearate, glyceryl dibehenate, glyceryl palmitostearate, hydrogenated vegetable oil, hydrogenated cottonseed oil, castor oil, mineral oil, polyethylene glycol (e.g., PEG 4000-8000), polyoxyethylene glycol, poloxamer, povidone, crospovidone, crosslinked sodium carboxymethylcellulose, alginic acid, casein, divinylbenzene methacrylate copolymer, docusate sodium, cyclodextrin (e.g., 2-hydroxypropyl-δ-cyclodextrin), polysorbate (e.g., polysorbate 80), cetrimide, TPGS (d-α-tocopheryl polyethylene glycol 1000 succinate), magnesium lauryl sulfate, sodium lauryl sulfate, polyethylene glycol ether, difatty acid ester of polyethylene glycol, or polyoxyalkylene sorbitan fatty acid ester (e.g., polyoxyethylene sorbitan ester) ), polyoxyethylene sorbitan fatty acid esters, sorbitan fatty acid esters (e.g., sorbitan fatty acid esters from fatty acids such as oleic acid, stearic acid or palmitic acid), mannitol, xylitol, sorbitol, maltose, lactose, lactose monohydrate or spray-dried lactose, sucrose, fructose, calcium phosphate, calcium hydrogen phosphate, tricalcium phosphate, calcium sulfate, dextrates, dextran, dextrin, D-glucose (dextrose), cellulose acetate, maltodextrin, simethicone, polydextrose, chitosan, gelatin, HPMC (hydroxypropyl methylcellulose), HPC (hydroxypropyl cellulose), hydroxyethyl cellulose, etc.

[0247] As will be appreciated by those skilled in the art, any diluent known in the art may be used according to the disclosure. In some embodiments of the disclosure, the diluent is water soluble. In some embodiments of the disclosure, the diluent is water insoluble. The term "diluent" as used herein includes, but is not limited to, water, saline, phosphate buffered saline (PBS), D-glucose, glycerol, ethanol, buffered sodium acetate or ammonium acetate solution, etc., and combinations thereof.

[0248] In some embodiments, the pharmaceutical composition of the present disclosure comprises at least one additional active ingredient. The term "active ingredient" as used herein refers to a therapeutically active compound, and any prodrug thereof, and pharmaceutically acceptable salts, hydrates, and solvates of the compound and prodrug. Additional active ingredients can be combined with the antibody of the present disclosure or its fragment, and can be administered separately or in the same pharmaceutical composition. Those skilled in the art can determine the amount of the additional active ingredient to be administered based on the therapy utilizing the antibody of the present disclosure or its fragment.

[0249] In some embodiments, the pharmaceutical composition is a human pharmaceutical composition.The term "human pharmaceutical composition" as used herein refers to a pharmaceutical composition for administration to humans.

[0250] The pharmaceutical compositions of the present disclosure are suitable for being applied to patients by any suitable means, including but not limited to those used for applying conventional drugs. The pharmaceutical compositions of the present disclosure can be applied using any suitable route considered by those skilled in the art, including but not limited to oral, intravenous ("IV") injection or infusion, intravesical, subcutaneous ("SC"), intramuscular ("IM"), intraperitoneal, intradermal, intraocular, inhalation (and intrapulmonary), intranasal, transdermal, epidermal, subdermal, topical, mucosal, nasal, ocular, skin impression into skin, intravaginal, intrauterine, intracervical and rectal. Such dosage forms should allow the antibodies of the present disclosure or their fragments to reach target cells. Other factors are well known in the art, including considerations such as toxicity and dosage forms that hinder compounds or compositions from exerting their effects. Techniques and formulations can generally be found in Remington: The Science and Practice of Pharmacy, 21st edition, Lippincott, Williams and Wilkins, Philadelphia, Pa., 2005.

[0251] In some embodiments, the pharmaceutical composition of the present disclosure is suitable for topical administration. The term "topical administration" as used herein refers to applying the antibody of the present disclosure or its fragment to the skin surface of the patient so that the antibody of the present disclosure or its fragment passes through the skin layer. Transdermal administration and transmucosal administration are also encompassed within the term topical administration. The term "transdermal" as used herein refers to an antibody of the present disclosure or its fragment passing through at least one skin layer of the patient. "Transmucosal" as used herein refers to an antibody of the present disclosure or its fragment passing through the mucous membrane of the patient. Unless otherwise specified or implied, the terms "topical administration", "transdermal administration" and "transmucosal administration" are used interchangeably herein.

[0252] A variety of topical delivery systems for delivering bioactive compounds to microorganisms in patients are well known in the art. Such systems include, but are not limited to, lotions, creams, gels, oils, ointments, solutions, suspensions, emulsions, etc., and suitable carriers in the art may be selected. In some embodiments, the pharmaceutical composition is administered in the form of a gel comprising a polyol.

[0253] Suitable carriers include, but are not limited to, vegetable or mineral oils, white petrolatum (e.g., white soft paraffin), branched chain fats or oils, animal fats, and high molecular weight alcohols (e.g., greater than C 12 ). In some embodiments, the carrier is selected so that the antibody or fragment thereof of the present disclosure is soluble. In some embodiments, emulsifiers, stabilizers, humectants and antioxidants and agents that impart color or fragrance (if necessary) may also be included. In some embodiments, organic solvents or co-solvents, such as ethanol or propanol, may be used in the pharmaceutical composition of the present disclosure. In some embodiments, the evaporation of the solvent will leave residues on the treated surface. In some embodiments, a penetrant suitable for the barrier to be penetrated is used. Such penetrants are generally known in the art and include but are not limited to bile salts and fusidic acid derivatives. In some embodiments, a detergent may be used to promote penetration. In some embodiments, a cream for topical application is formulated with a mixture of mineral oil, self-emulsifying beeswax and water, wherein the antibody or fragment thereof of the present disclosure dissolved in a small amount of solvent (e.g., oil) is mixed in the mixture. The specific topical delivery system used depends on the intended site of application.

[0254] In some embodiments, other materials can also be added to the topical pharmaceutical composition of the present disclosure to have a moisturizing effect and improve the consistency of the pharmaceutical composition. The example of this compound includes but is not limited to hexadecyl ester wax, stearyl alcohol, cetyl alcohol, glycerol, methylparaben, propylparaben, quaternary ammonium salt-15, humectant, volatile methylsiloxane fluid and polydiorganosiloxane-polyoxyalkylene. For example, referring to U.S. Patent number 5,153,230 and 4,421,769. In some embodiments, if it is desired that the pharmaceutical composition has an additional cleaning effect, chemicals such as sodium lauryl sulfate or carboxylic acid metal salts etc. can be added.

[0255] In some embodiments, a variety of non-volatile emollients can be used for pharmaceutical compositions of the present disclosure. Non-limiting examples of such non-volatile emollients are listed in: McCutcheon's, Volume 2 Functional Materials, North American Edition, (1992), pages 137-168; and CTFA Cosmetic Ingredient Handbook, 2nd edition (1992) lists skin conditioning agents at pages 572-575 and skin protectants at page 580. In some embodiments, non-volatile emollients include silicones, hydrocarbons, esters, and mixtures thereof. In some embodiments, esters include esters of monofunctional and difunctional fatty acids esterified with alcohols and polyols (i.e., alcohols with two or more hydroxyls). In some embodiments, long-chain esters of long-chain fatty acids (i.e., C10-40 fatty acids esterified with C10-40 fatty alcohols) are used in pharmaceutical compositions of the present disclosure. Non-limiting examples of esters that can be used in the pharmaceutical compositions of the present disclosure include, but are not limited to, esters selected from the group consisting of diisopropyl adipate, isopropyl myristate, isopropyl palmitate, myristyl propionate, ethylene glycol distearate, 2-ethylhexyl palmitate, isodecyl neopentanoate, C12-15 alcohol benzoate, di-2-ethylhexyl maleate, hexadecyl palmitate, myristyl myristate, octadecyl stearate, hexadecyl stearate, behenyl behenrate, and mixtures thereof.

[0256] Examples of silicone emollients that can be used in the pharmaceutical compositions of the present disclosure include, but are not limited to, polyalkylsiloxanes, cyclic polyalkylsiloxanes, and polyalkylarylsiloxanes. Suitable commercially available polyalkylsiloxanes include polydimethylsiloxanes, also known as dimethicones, non-limiting examples of which include VICASIL sold by General Electric Company. TM Series and DOW CORNING sold by Dow Corning TM 200 series. Commercially available polyalkylsiloxanes include cyclomethicone (DOW CORNING TM 244 Fluid), DOW CORNING TM 344 Fluid, Down Corning TM 245 Fluid and Down Corning TM 345) etc. Suitable commercially available trimethylsiloxysilicates are used as a mixture with polydimethylsiloxane, such as DOW CORNING TM593 fluid. Also included in the pharmaceutical composition of the present disclosure is polydimethylsiloxane, which is a hydroxyl-terminated dimethyl silicone oil. Suitable commercially available polydimethylsiloxane is usually sold as a mixture with polydimethylsiloxane or cyclomethicone (e.g., DOW CORNING TM 1401, 1402, and 1403 fluids). Suitable commercially available polyalkylaryl siloxanes include SF1075 methyl phenyl fluid (sold by General Electric Company) and 556 Cosmetic Grade Phenyl Trimethicone Fluid (sold by Dow Corning Corporation).

[0257] Hydrocarbons suitable for use in pharmaceutical compositions of the present disclosure include, but are not limited to, straight and branched hydrocarbons having from about 10 to about 30 carbon atoms. In some embodiments, straight and branched hydrocarbons have from about 12 to about 24 carbon atoms. In some embodiments, straight and branched hydrocarbons have from about 16 to about 22 carbon atoms. Non-limiting examples of such hydrocarbon materials include dodecane, squalane, cholesterol, 5-hydrogenated polyisobutene, docosane (i.e., C22 hydrocarbons), hexadecane, and isohexadecane (a commercial hydrocarbon available from Prespers of South Plainsfield, New Jersey as PERMETHYL TM 101A Sales) etc.

[0258] In some embodiments, topical pharmaceutical compositions of the present disclosure include propylene glycol. In some embodiments, propylene glycol plays a role as a surfactant and contributes to the penetration, contact and absorption of the antibodies of the present disclosure or their fragments. In some embodiments, propylene glycol is used as a preservative. In some embodiments, pharmaceutical compositions of the present disclosure include nonionic surfactants, such as polysorbates. Such surfactants provide pharmaceutical compositions of the present disclosure with better surface contact with mucosa (e.g., vaginal mucosa) by further reducing surface tension.

[0259] The topical pharmaceutical composition of the present disclosure may also be optionally formulated with a lipophilic phase, such as with an emulsion and a liposome dispersion. In some embodiments, the liposome formulation can prolong the circulation time of the antibody of the present disclosure or its fragment, increase the permeability of the antibody of the present disclosure or its fragment, and improve the overall efficacy of the antibody of the present disclosure or its fragment as a drug. In some embodiments, the antibody of the present disclosure or its fragment can be combined with a lipid, a cationic lipid or an anionic lipid. In some embodiments, the resulting emulsion or liposome suspension can effectively increase the in vivo active half-life of the pharmaceutical composition of the present disclosure. Examples of anionic lipids suitable for pharmaceutical compositions of the present disclosure include, but are not limited to, cardiolipin, dimyristoyl, dipalmitoyl, dioleoylphosphatidylcholine, phosphatidylglycerol, palmitoyloleoylphosphatidylcholine, phosphatidylglycerol, phosphatidic acid, lysophosphatidic acid, phosphatidylserine, phosphatidylinositol and anionic forms of cholesterol.

[0260] In some embodiments, the antibody of the present disclosure or its fragment is incorporated into liposome. In some embodiments, neutral lipid, cholesterol and / or polyethylene glycol (PEG) are used in such liposome. In some embodiments, liposome composition is made up of partially hydrogenated soybean phosphatidylcholine (PHSC), cholesterol, methoxy-terminated polyethylene glycol (mPEG) and / or distearoylphosphatidylethanolamine (DSPE). Liposome can be prepared according to any suitable method known in the art.

[0261] In some embodiments, topical application is carried out by nasal spray or suppository (rectal or vaginal). Suppository is prepared by mixing the antibody of the present disclosure or its fragment with a lipid carrier (e.g., cocoa butter, cocoa butter, glycerine, gelatin, polyoxyethylene glycol, etc.). In some embodiments, topical application includes transdermal patches or dressings, such as bandages impregnated with the antibody of the present disclosure or its fragment and optionally one or more carriers, excipients or diluents known in the art. In some embodiments, such dressings include but are not limited to semipermeable membranes, foams, hydrocolloids and calcium alginate swabs. In some embodiments, dosage administration will be continuous rather than intermittent throughout the dosage regimen.

[0262] In some embodiments, the pharmaceutical composition of the present disclosure is suitable for oral administration. The term "oral administration" used herein refers to the administration of the antibody of the present disclosure or its fragment to the patient's mouth for ingestion into the gastrointestinal tract. In some embodiments, the pharmaceutical composition of the present disclosure can be formulated into conventional oral dosage forms, including but not limited to capsules, tablets, powders and liquid preparations, such as suspensions, solutions, elixirs, syrups, concentrated drops, etc. In some embodiments, the antibody of the present disclosure or its fragment can be combined with a solid excipient, optionally grinding the resulting mixture, and optionally processing the particle mixture after adding a suitable auxiliary agent (if necessary), to obtain, for example, tablets, coated tablets, hard capsules, soft capsules, solutions (e.g., aqueous solutions, alcoholic solutions or oily solutions), etc. In some embodiments, excipients suitable for oral pharmaceutical compositions of the present disclosure include, but are not limited to, fillers, such as sugars, including lactose, glucose, sucrose, mannitol or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose (CMC) and / or polyvinyl pyrrolidone (PVP or povidone); and oily excipients, including vegetable oils and animal oils, such as sunflower oil, olive oil or cod liver oil. In some embodiments, oral pharmaceutical compositions of the present disclosure may also contain disintegrants, such as cross-linked polyvinyl pyrrolidone, agar or alginic acid, or salts thereof, such as sodium alginate; lubricants, such as talc or magnesium stearate; plasticizers, such as glycerol or sorbitol; sweeteners, such as sucrose, fructose, lactose or aspartame; natural or artificial flavorings, such as mint, wintergreen oil or cherry flavoring; or dyes or pigments, which can be used to identify or characterize different doses or combinations. In some embodiments, the oral pharmaceutical compositions of the present disclosure may also contain dragee cores with suitable coatings. In some embodiments, concentrated sugar solutions may be used, which may optionally contain, for example, gum arabic, talc, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol, titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.

[0263] In some embodiments, pharmaceutical compositions of the present disclosure that can be used orally include, but are not limited to, push-in capsules made of gelatin ("gel capsules"), and soft sealed capsules made of gelatin and a plasticizer (e.g., glycerol or sorbitol). In some embodiments, the push-in capsules may contain a mixture of an antibody or fragment thereof of the present disclosure with a filler (e.g., lactose), a binder (e.g., starch), and / or a lubricant (e.g., talc or magnesium stearate), and optionally a stabilizer. In some embodiments including soft capsules, the antibody or fragment thereof may be dissolved or suspended in a suitable liquid, such as a fatty oil, liquid paraffin, liquid polyethylene glycol, and the like.

[0264] In some embodiments, the pharmaceutical composition of the present disclosure is suitable for inhalation administration. The term "inhalation administration" used herein refers to delivering the antibody or its fragment of the present disclosure through the patient's nose or mouth during the inhalation process, and allowing the antibody or its fragment to pass through the patient's lung wall. In some embodiments, the pharmaceutical composition of the present disclosure suitable for inhalation administration can be formulated as a dry powder or a suitable solution, suspension or aerosol. In some embodiments, powders and solutions can be formulated with suitable additives known in the art. In some embodiments, powders can include a suitable powder base, such as lactose or starch. In some embodiments, solutions can include propylene glycol, sterile water, ethanol, sodium chloride and other additives, such as acids, alkalis and buffer salts. In some embodiments, such solutions or suspensions can be administered by inhalation such as sprays, pumps, sprayers, nebulizers, etc. In some embodiments, the pharmaceutical compositions of the present disclosure suitable for inhaled administration may also be used in combination with other inhaled therapies, including but not limited to corticosteroids such as fluticasone propionate, beclomethasone propionate, triamcinolone acetonide, budesonide, and mometasone furoate; beta agonists such as albuterol, salmeterol, and formoterol; anticholinergic agents such as ipratropium bromide or tiotropium bromide; vasodilators such as treprostinil and iloprost; enzymes such as DNAase; therapeutic proteins; immunoglobulin antibodies; oligonucleotides such as single-stranded or double-stranded DNA or RNA, siRNA; antibiotics such as tobramycin; muscarinic receptor antagonists; leukotriene antagonists; cytokine antagonists; protease inhibitors; sodium cromoglycate; sodium nedocrine; and sodium cromoglycate.

[0265] In some embodiments, the pharmaceutical compositions of the present disclosure are suitable for intravesical administration. As used herein, the term "intravesical administration" refers to delivering the antibodies of the present disclosure or fragments thereof directly into the bladder of a patient. In some embodiments, the pharmaceutical compositions are administered via a catheter. In some embodiments, the catheter is a urethral catheter.

[0266] In some embodiments, the pharmaceutical compositions of the present disclosure are suitable for parenteral administration. As used herein, the term "parenteral administration" refers to the injection or infusion of the antibody or fragment thereof of the present disclosure into a patient, including but not limited to intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, intracerebrospinal and intrasternal injection and infusion. In some embodiments, the pharmaceutical compositions of the present disclosure suitable for parenteral administration can be formulated in a sterile liquid solution, including but not limited to a physiologically compatible buffer or solution, such as a saline solution, Hank's solution or Ringer's solution. In some embodiments, the pharmaceutical compositions of the present disclosure suitable for parenteral administration can be prepared as a dispersant in a non-aqueous solution, such as glycerol, propylene glycol, ethanol, liquid polyethylene glycol, triacetin, vegetable oil, etc. In some embodiments, the solution may also contain preservatives, such as methylparaben, propylparaben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In addition, pharmaceutical compositions of the present disclosure suitable for parenteral administration may be formulated in solid form, including, for example, lyophilized form, and re-dissolved or suspended prior to use. In some embodiments, the pharmaceutical composition is administered by needle. For injection, the carrier may be a liquid, such as sterile pyrogen-free water, pyrogen-free phosphate buffered saline solution, bacteriostatic water, or CREMOPHOR (BASF, Parsippany, NJ.) For other methods of administration, the carrier may be either a solid or a liquid.

[0267] The present disclosure also provides a kit. In some embodiments, the kit comprises an antibody or fragment thereof according to the present disclosure. As used herein, the term "kit" refers to any article, such as packaging, container, etc., which contains an antibody or fragment thereof according to the present disclosure. In some embodiments, an antibody or fragment thereof according to the present disclosure is packaged in a vial, bottle, tube, flask or patch, which may be further packaged in a box, envelope, bag, etc. In some embodiments, an antibody or fragment thereof according to the present disclosure has been approved by the U.S. Food and Drug Administration or similar regulatory agencies in jurisdictions or regions outside the United States or the United States and can be applied to patients. In some embodiments, the kit includes written instructions for use and / or indicates that the antibody or fragment thereof according to the present disclosure is suitable for or approved for application to other indications of the patient. In some embodiments, the antibody or fragment thereof of the present disclosure is packaged in unit dose or single unit dose form, such as single unit dose pills, capsules, etc. In some embodiments, the kit includes a dispenser.

[0268] The present disclosure also provides the use of the antibody or fragment thereof of the present disclosure for preparing a medicament. The term "medicament" as used herein refers to a pharmaceutical composition according to the present disclosure. In some embodiments, the pharmaceutical composition is contained in any article, such as a package, a container, etc.

[0269] As one aspect, a method of depleting, inhibiting, or depleting and inhibiting pDC in a patient comprises delivering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising BDCA2 and depletes, inhibits, or depletes and inhibits pDC, thereby depleting, inhibiting, or depleting and inhibiting pDC. The antibody or fragment thereof is as described above. In addition, the antibody or fragment thereof can be formulated in a pharmaceutical composition as described above.

[0270] The term "depletion" used herein with respect to pDC refers to a measurable reduction in the number of pDCs in a patient or sample. The reduction may be at least about 10%, for example at least about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more. In certain embodiments, the term refers to a reduction in the number of pDCs in a patient or sample to an amount below the detectable limit. In some embodiments, pDCs are consumed by at least about 50%, for example at least about 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more, compared to patients who have not received antibodies or fragments thereof. In other embodiments, the disclosure provides a method for reducing the number of pDCs and / or consuming pDCs in a sample (e.g., a mixed cell population) in vitro or in vitro, the method comprising delivering an effective amount of an antibody or fragment thereof that specifically binds to BDCA2 and reduces the number of pDCs and / or consumes pDCs to the sample, thereby reducing the number of pDCs and / or consuming pDCs. In some embodiments, BDCA2 is human BDCA2.

[0271] The term "inhibition" used herein with respect to pDC refers to slowing down or eliminating certain activities of pDC, such as cell survival, pDC activation (e.g., the production of type 1 interferon), and any other activity of the cell. Various methods of quantitative and / or qualitative cell inhibition are within the capabilities of those of ordinary skill in the art.

[0272] In some embodiments, a method of treating a pDC-associated disorder in a patient in need thereof comprises delivering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen 2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDCs, thereby treating the disorder. The antibody or fragment thereof is as described above. In addition, the antibody or fragment thereof can be formulated in a pharmaceutical composition as described above.

[0273] The present disclosure provides a method of treatment. The terms "treatment", "therapy" and similar terms used herein refer to the administration of an effective amount of an antibody or fragment thereof or a pharmaceutical composition of the present disclosure to prevent, alleviate or improve one or more symptoms of a disease or condition (i.e., an indication) and / or to prolong the survival of the treated patient. In some embodiments, "treatment", "therapy" and similar terms also include, but are not limited to, reducing or eliminating an infection in a patient.

[0274] When implementing the method of the present disclosure, an effective amount of the antibody or fragment thereof of the present disclosure is administered to a patient in need. As used herein, the term "effective amount" refers to the amount of the antibody or fragment thereof or pharmaceutical composition of the present disclosure in the context of administration, which, when administered to a patient, is sufficient to prevent, alleviate or improve one or more symptoms in a disease or condition (i.e., an indication) and / or prolong the survival of the treated patient. Such doses should not cause adverse reactions in the treated patient, or cause very few adverse reactions. Similarly, such doses should not cause toxic effects in the treated patient, or cause very few toxic effects. It will be understood by those skilled in the art that the amount of the antibody or fragment thereof or pharmaceutical composition of the present disclosure will vary according to many factors, including but not limited to the activity of the antibody or fragment thereof of the present disclosure (e.g., the in vitro activity of the compound of the present disclosure on the target, or the in vivo activity in an animal model), the pharmacokinetic results in an animal model (e.g., biological half-life or bioavailability), the type of the treated patient, the age, size, weight and general physical condition of the patient, the disease associated with the patient, and the dosing regimen used in the treatment. Those skilled in the art will appreciate that the therapeutic effect need not be complete or curative, as long as some benefit is provided to the patient.

[0275] In some embodiments of the present disclosure, the effective amount of the antibody or fragment of the present disclosure to be delivered to a patient in need thereof can be quantified by determining the micrograms of the antibody or fragment of the present disclosure per kilogram of patient body weight. In some embodiments, the amount of the antibody or fragment of the present disclosure administered to the patient is about 0.1 to about 1000 milligrams (mg) of the antibody or fragment of the present disclosure per kilogram (kg) of patient body weight. In some embodiments, the amount of the antibody or fragment of the present disclosure administered to the patient is about 0.1 to about 500 mg of the antibody or fragment of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment of the present disclosure administered to the patient is about 0.1 to about 300 mg of the antibody or fragment of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment of the present disclosure administered to the patient is about 0.1 to about 200 mg of the antibody or fragment of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment of the present disclosure administered to the patient is about 0.1 to about 100 mg of the antibody or fragment of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment thereof of the present disclosure administered to the patient is about 0.1 to about 80 mg of the antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment thereof of the present disclosure administered to the patient is about 0.1 to about 50 mg of the antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment thereof of the present disclosure administered to the patient is about 0.1 to about 20 mg of the antibody or fragment thereof of the present disclosure per kg of patient body weight. In some embodiments, the amount of the antibody or fragment thereof of the present disclosure administered to the patient is about 0.1 to about 10 mg of the antibody or fragment thereof of the present disclosure per kg of patient body weight. As will be appreciated by those of ordinary skill in the art, multiple doses may be used.

[0276] In some embodiments of the present disclosure, the antibodies or fragments thereof of the present disclosure are administered as a multiple dose regimen. The term "multiple dose regimen" as used herein refers to a treatment period of more than one day. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 2 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 3 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 4 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 5 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 6 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 7 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about 14 days. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about one month. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about two months. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about three months. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about four months. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about five months. In some embodiments of the present disclosure, the multiple dose regimen is a period of up to about six months. Other time periods may also be used herein.

[0277] In some embodiments of the present disclosure, the antibodies or fragments thereof of the present disclosure are administered as part of a chronic treatment regimen. As used herein, the term "chronic treatment regimen" refers to treatment with the antibodies or fragments thereof of the present disclosure over a longer period of time during the patient's lifetime. In some embodiments, chronic treatment is lifelong treatment.

[0278] In some embodiments of the present disclosure, the antibodies of the present disclosure or fragments thereof are administered as a single dose. In some embodiments of the present disclosure, the antibodies of the present disclosure or fragments thereof are administered as a single unit dose. As used herein, the term "unit dose" is a predetermined amount of an antibody of the present disclosure or fragments thereof. The amount of an antibody of the present disclosure or fragments thereof is generally equal to the dosage of the antibody of the present disclosure or fragments thereof administered to the patient, or a convenient fraction of the dosage, such as half or one-third of the dosage. According to the methods of the present disclosure, the terms "single dose" and "single unit dose" include embodiments in which the antibodies of the present disclosure or fragments thereof can be administered as a single application and as multiple applications.

[0279] In some embodiments, the antibodies or fragments thereof of the present invention may also be used in combination with one or more additional active ingredients for the treatment of the same disease or condition. In some embodiments, such combined use includes administering the antibodies or fragments thereof of the present invention and one or more additional active ingredients at different times, or co-administering the antibodies or fragments thereof of the present invention and one or more additional active ingredients. In some embodiments, the dosage of the antibodies or fragments thereof of the present invention or one or more additional active ingredients used in combination may be modified by methods well known to those of ordinary skill in the art, for example, reducing the dosage of the antibodies or fragments thereof of the present invention or one or more additional active ingredients used alone. In some embodiments, co-administration includes administering the antibodies or fragments thereof of the present invention and additional active ingredients simultaneously in the same dosage form, administering the antibodies or fragments thereof of the present invention and additional active ingredients simultaneously in separate dosage forms, and administering the antibodies or fragments thereof of the present invention and additional active ingredients separately.

[0280] It should be understood that combined use includes use with one or more additional active ingredients or other medical procedures, where one or more additional active ingredients or other medical procedures can be administered at a different time than the antibody or fragment thereof or pharmaceutical composition of the present disclosure (e.g., within a short period of time, such as within a few hours (e.g., about 1, 2, 3, 4-24 hours, etc.), or within a longer period of time (e.g., about 1-2 days, 2-4 days, 4-7 days, 1-4 weeks, etc.)), or administered simultaneously with the antibody or fragment thereof or pharmaceutical composition of the present disclosure. Combined use also includes use with one or more additional active ingredients or other medical procedures (e.g., surgery) that are administered once or infrequently, where the antibody or fragment thereof or pharmaceutical composition of the present disclosure is administered within a short period of time or longer before or after the administration of one or more additional active ingredients or the completion of the other medical procedure.

[0281] In some embodiments, the present disclosure provides antibodies or fragments thereof or pharmaceutical compositions of the present disclosure and one or more additional active ingredients delivered by different routes of administration or by the same route of administration. In some embodiments, the combined use for any route of administration includes delivering antibodies or fragments thereof or pharmaceutical compositions of the present disclosure and one or more additional active ingredients in any pharmaceutical composition together with the same route of administration, including pharmaceutical compositions in which two compounds are chemically connected so that these compounds maintain their therapeutic activity when administered. In some embodiments, one or more additional active ingredients can be co-administered with antibodies or fragments thereof or pharmaceutical compositions of the present disclosure. In some embodiments, the combined use by co-administration includes administering a preparation of a co-preparation or chemically connected compound, or administering two or more compounds with separate preparations by the same or different routes within a short period of time (e.g., within about one hour, 2 hours, 3 hours, up to about 24 hours, etc.). In some embodiments, the co-administration of separate preparations includes co-administration delivered by a device, for example, the same inhalation device, the same syringe, etc., or administering each other by separate devices within a short period of time. In some embodiments, co-formulations of the antibodies or fragments thereof or pharmaceutical compositions of the present disclosure with one or more additional active ingredients delivered by the same route include co-formulations of these materials so that they can be administered by one device, separate compounds combined in one formulation, or compounds modified so that the compounds are chemically linked but still retain their biological activity. In some embodiments, such chemically linked compounds may have a bond that is substantially maintained in vivo, or may be broken in vivo, thereby separating the two active ingredients.

[0282] As used herein, the phrase "disease associated with pDC" refers to any disease, disorder or condition in which pDC plays a role in the cause, side effect, symptom or other aspects of the disease, disorder or condition. Examples of such disorders include, but are not limited to, autoimmune diseases, inflammatory disorders, respiratory disorders, skin disorders, cancer, and combinations of two or more thereof.

[0283] As used herein, the term "infectious disease" refers to any disease associated with infection by an infectious agent. Examples of infectious agents include, but are not limited to, viruses and microorganisms. Viruses include, but are not limited to: Hepadnaviridae, including hepatitis A, B, C, D, E, F, G, etc.; Flaviviridae, including human hepatitis C virus (HCV), yellow fever virus, and dengue virus; Retroviridae, including human immunodeficiency virus (HIV) and human T-lymphotropic virus (HITLV1 and HTLV2); Herpesviridae, including herpes simplex virus (HSV-1 and HSV-2), Epstein-Barr virus (EBV-1 and EBV-2); V), cytomegalovirus, varicella-zoster virus (VZV), human herpesvirus 6 (HHV-6), human herpesvirus 8 (HHV-8), and herpes B virus; Papovaviridae, including human papillomavirus; Rhabdoviridae, including rabies virus; Paramyxoviridae, including respiratory syncytial virus; Reoviridae, including rotavirus; Bunyaviridae, including hantavirus; Filoviridae, including Ebola virus; Adenoviridae; Parvoviridae, including parvovirus B-19; Arenaviridae, including Lassa virus; Orthomyxoviridae, including influenza virus; Poxviridae, including Orf virus, molluscum contagiosum virus, variola virus, and monkeypox virus; Togaviridae, including Venezuelan equine encephalitis virus; Coronaviridae, including coronaviruses, such as severe acute respiratory syndrome (SARS) virus (e.g., SARS-CoV-2); and Picornaviridae, including poliovirus; Rhinovirus; Orbivirus; Picornavirus; Encephalomyocarditis virus (EMV); parainfluenza virus, adenovirus, coxsackievirus, echovirus, measles virus, rubella virus, human papillomavirus, canine distemper virus, canine infectious hepatitis virus, feline calicivirus, feline rhinotracheitis virus, TGE virus (swine), foot-and-mouth disease virus, simian virus 5, human parainfluenza virus type 2, human subpneumovirus, enterovirus, and any other pathogenic virus now known or later identified (see, e.g., Fundamental Virology, Fields et al., 3rd ed., Lippincott-Raven, New York, 1996, the entire contents of which are incorporated herein by reference for its teachings regarding pathogenic viruses).

[0284] Pathogenic microorganisms include, but are not limited to, Rickettsia, Chlamydia, Mycobacteria, Clostridia, Corynebacteria, Mycoplasma, Ureaplasma, Legionella, Shigella, Salmonella, pathogenic Escherichia coli species, Bordatella, Neisseria, Treponema, Bacillus, Haemophilus, Moraxella, Vibrio, Staphylococcus spp., Streptococcus spp., Campylobacter spp., Borrelia spp.), Leptospira spp., Erlichia spp., Klebsiella spp., Pseudomonas spp., Helicobacter spp., and any other pathogenic microorganism now known or later identified (e.g., see Microbiology, Davis et al., 4th ed., Lippincott, New York, 1990, the entire contents of which are incorporated herein by reference for teachings regarding pathogenic microorganisms).Specific examples of microorganisms include, but are not limited to, Helicobacter pylori, Chlamydia pneumoniae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Treponemapallidum, Bacillus anthracis, Salmonella typhi, Vibrio cholera. pestis (Pasteurella pestis), (Yersinia pestis), Pseudomonas aeruginosa, Campylobacter jejuni, Clostridium dificile, Clostridium botulinum, Mycobacterium tuberculosis, Borrelia burgdorferi, Haemophilus ducreyi, Corynebacterium diphtheria, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenza, and enterotoxic Escherichia coli.

[0285] As used herein, the term "autoimmune disorder" refers to any disorder associated with an autoimmune response. Examples include, but are not limited to, diabetes, psoriasis of the skin, hyperthyroidism, autoimmune adrenal insufficiency, hemolytic anemia, rheumatic myocarditis, systemic lupus erythematosus, cutaneous lupus erythematosus, psoriatic arthritis, Sjögren's syndrome polymyositis, myositis, multiple sclerosis, Crohn's disease, ulcerative colitis, lupus, inflammatory bowel syndrome, dermatomyositis, dermatitis, atopic dermatitis, scleroderma, polymyositis, psoriasis, alopecia areata, rheumatoid arthritis, graft versus host disease, interferon disease, and irritable bowel syndrome.

[0286] As used herein, the term "inflammatory disorder" refers to a disorder in which the tissues of an organism react to certain stimuli, including autoimmune disorders, commonly referred to as "inflammation." Symptoms of inflammation include swelling, pain, heat, redness, and loss of tissue function. Exemplary inflammatory disorders include neurodegenerative diseases; diabetic nephropathy and retinopathy; protein wasting, muscle fatigue or inflammation; infectious diseases; various cardiovascular diseases or disorders, including atherosclerosis; neurodegenerative diseases, such as Alzheimer's disease; infectious diseases, such as myocarditis, cardiomyopathy, acute endocarditis, pericarditis; systemic inflammatory response syndrome (SIRS) / sepsis; adult respiratory distress syndrome (ARDS); asthma; rheumatoid arthritis; osteoarthritis; juvenile arthritis; dry eye syndrome; systemic lupus erythematosus; airway hyperresponsiveness (AHR); bronchial hyperresponsiveness; chronic obstructive pulmonary disease (COPD); congestive heart failure (CHF); inflammatory complications of diabetes; metabolic syndrome; ankylosing spondylitis; end-stage renal disease (ESRD); hepatitis; liver fibrosis; pulmonary fibrosis; organ transplant rejection; acute kidney injury; and diseases associated with aging.

[0287] The term "respiratory disorder" as used herein refers to a pathological condition that affects organs and tissues, resulting in difficulty in gas exchange in air-breathing animals. Respiratory disorders include disorders of the respiratory tract, including disorders of the trachea, bronchi, bronchioles, alveoli, pleura, chest cavity, nerves, and respiratory muscles. Respiratory diseases range from mild self-limiting diseases (e.g., the common cold, influenza, and pharyngitis) to life-threatening diseases (e.g., bacterial pneumonia, pulmonary embolism, tuberculosis, acute or chronic asthma, pulmonary fibrosis, lung cancer, and severe acute respiratory syndrome (e.g., COVID-19)). Respiratory diseases can be classified in a variety of different ways, including classification according to the organs or tissues involved, classification according to the type and pattern of related signs and symptoms, or classification according to the cause of the disease.

[0288] As used herein, the term "skin disorder" refers to any medical condition that affects a patient's integumentary system - the integumentary system is an organ system surrounding the body, including the skin, hair, nails, and associated muscles and glands, whose primary function is to serve as a barrier against the external environment. Exemplary skin disorders include, but are not limited to, scleroderma, psoriasis, dermatomyositis, and atopic dermatitis.

[0289] As used herein, the term "cancer" refers to any abnormal growth of cells, whether benign or malignant. Examples include, but are not limited to, breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head and neck cancer, breast cancer, ovarian cancer, lung cancer, small cell lung cancer, Wilms' tumor, cervical cancer, testicular cancer, bladder cancer, pancreatic cancer, stomach cancer, colon cancer, prostate cancer, genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia Disease, blastic plasmacytoid dendritic cell neoplasm ("BPDCN" or "pDC leukemia"), chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia ("AML"), chronic myeloid leukemia, acute myeloid leukemia, senescence-associated hairy cell leukemia, chronic myelomonocytic leukemia ("CMML"), myelodysplastic syndrome, chronic myelofibrosis, multiple myeloma, neuroblastoma, rhabdomyosarcoma, Kaposi's sarcoma, Waldenstrom's macroglobulinemia, diffuse large B-cell lymphoma (DLBCL) (L265P), polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, essential macroglobulinemia and retinoblastoma. In some embodiments, the cancer is selected from a neoplastic cancer.

[0290] In addition to the aspects and embodiments described and provided elsewhere in this disclosure, the following non-limiting embodiments are also contemplated.

[0291] 1. An antibody or a fragment thereof, comprising:

[0292] a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0293] A light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0294] 2. The antibody or fragment thereof according to item 1, wherein the antibody or fragment thereof binds to the target protein with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binds to the epitope having SEQ ID NO:11.

[0295] 3. The antibody or fragment thereof according to item 1 or item 2, wherein the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0296] 4. The antibody or fragment thereof according to items 1 to 3, wherein the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0297] 5. The antibody or fragment thereof according to items 1 to 4, wherein:

[0298] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0299] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0300] 6. The antibody or fragment thereof according to items 1 to 4, wherein:

[0301] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0302] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0303] 7. The antibody or fragment thereof according to items 1 to 4, wherein:

[0304] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0305] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0306] 8. The antibody or fragment thereof according to items 1 to 4, wherein:

[0307] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0308] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0309] 9. The antibody or fragment thereof according to items 1 to 4, wherein:

[0310] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0311] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0312] 10. The antibody or fragment thereof according to items 1 to 4, wherein:

[0313] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0314] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0315] 11. The antibody or fragment thereof according to items 1 to 4, wherein:

[0316] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0317] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0318] 12. The antibody or fragment thereof according to items 1 to 4, wherein:

[0319] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0320] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0321] 13. The antibody or fragment thereof according to items 1 to 4, wherein:

[0322] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0323] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0324] 14. The antibody or fragment thereof according to items 1 to 4, wherein:

[0325] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0326] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0327] 15. The antibody or fragment thereof according to item 1 to 5 or 9, wherein:

[0328] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0329] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0330] 16. The antibody or fragment thereof according to item 1 to 4, 6 or 9, wherein:

[0331] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0332] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0333] 17. The antibody or fragment thereof according to item 1 to 4, 7 or 9, wherein:

[0334] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0335] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0336] 18. The antibody or fragment thereof according to item 1 to 4, 8 or 9, wherein:

[0337] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0338] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0339] 19. The antibody or fragment thereof according to item 1 to 5 or 10, wherein:

[0340] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0341] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0342] 20. The antibody or fragment thereof according to item 1 to 5 or 11, wherein:

[0343] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0344] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0345] 21. The antibody or fragment thereof according to item 1 to 5 or 12, wherein:

[0346] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0347] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0348] 22. The antibody or fragment thereof according to item 1 to 5 or 13, wherein:

[0349] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0350] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0351] 23. The antibody or fragment thereof according to item 1 to 5 or 14, wherein:

[0352] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0353] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0354] 24. The antibody or fragment thereof according to item 1 to 4, 6 or 10, wherein:

[0355] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0356] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0357] 25. The antibody or fragment thereof according to item 1 to 4, 7 or 10, wherein:

[0358] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0359] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0360] 26. The antibody or fragment thereof according to item 1 to 4, 8 or 10, wherein:

[0361] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0362] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0363] 27. The antibody or fragment thereof according to item 1 to 4, 6 or 11, wherein:

[0364] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0365] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0366] 28. The antibody or fragment thereof according to item 1 to 4, 7 or 11, wherein:

[0367] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0368] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0369] 29. The antibody or fragment thereof according to item 1 to 4, 8 or 11, wherein:

[0370] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0371] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0372] 30. The antibody or fragment thereof according to item 1 to 4, 6 or 12, wherein:

[0373] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0374] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0375] 31. The antibody or fragment thereof according to item 1 to 4, 7 or 12, wherein:

[0376] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0377] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0378] 32. The antibody or fragment thereof according to item 1 to 4, 8 or 12, wherein:

[0379] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 4, or a sequence at least about 90% identical to SEQ ID NO: 4; and

[0380] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0381] 33. The antibody or fragment thereof according to item 1 to 4, 6 or 13, wherein:

[0382] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0383] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0384] 34. The antibody or fragment thereof according to item 1 to 4, 7 or 13, wherein:

[0385] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0386] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0387] 35. The antibody or fragment thereof according to item 1 to 4, 8 or 13, wherein:

[0388] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0389] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0390] 36. The antibody or fragment thereof according to item 1 to 4, 6 or 14, wherein:

[0391] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0392] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0393] 37. The antibody or fragment thereof according to item 1 to 4, 7 or 14, wherein:

[0394] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0395] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0396] 38. The antibody or fragment thereof according to item 1 to 4, 8 or 14, wherein:

[0397] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0398] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0399] 39. A method for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient in need thereof, the method comprising administering to the patient an effective amount of an antibody or fragment thereof that specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDCs, thereby depleting, inhibiting, or depleting and inhibiting pDCs; wherein the antibody or fragment thereof comprises:

[0400] a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0401] A light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0402] 40. The method according to item 39, wherein the epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof binds to the epitope with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binding epitope.

[0403] 41. The method according to item 39 or item 40, wherein the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0404] 42. The method according to items 39 to 41, wherein the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0405] 43. The method according to items 39 to 42, wherein the BDCA2 is human BDCA2.

[0406] 44. A method for treating a disorder associated with plasmacytoid dendritic cells (pDCs) in a patient in need thereof, the method comprising:

[0407] An effective amount of an antibody or fragment thereof is administered to the patient, wherein the antibody or fragment thereof specifically binds to an epitope comprising blood dendritic cell antigen-2 (BDCA2) and depletes, inhibits, or depletes and inhibits pDC, thereby treating the disorder; wherein the antibody or fragment thereof comprises:

[0408] a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0409] A light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0410] 45. The method according to item 44, wherein the disease is selected from infectious diseases, autoimmune diseases, inflammatory disorders, respiratory disorders, skin disorders, cancer and combinations of two or more thereof.

[0411] 46. ​​The method according to item 45, wherein the disease is an infectious disease associated with infection with an infectious agent.

[0412] 47. The method according to item 46, wherein the infectious agent is one or more viruses or one or more microorganisms.

[0413] 48. The method of claim 47, wherein the infectious agent is a virus selected from the group consisting of: Hepadnaviridae; Flaviviridae; Retroviridae; Herpesviridae; Papovaviridae; Rhabdoviridae; Paramyxoviridae; Reoviridae; Bunyaviridae; Filoviridae; Adenoviridae; Parvoviridae; Arenaviridae; Orthomyxoviridae; Poxviridae; Togaviridae; Coronaviridae; Picornaviridae; Rhinovirus; Orbivirus; Picodnavirus; Encephalomyocarditis virus (EMV); Parainfluenza virus; Adenovirus; Coxsackievirus; Echovirus; Measles virus; Rubella virus; Human papillomavirus; Canine distemper virus; Canine infectious hepatitis virus; Feline calicivirus; Feline rhinotracheitis virus; TGE virus (swine); Foot-and-mouth disease virus; Simian virus 5; Human parainfluenza virus type 2; Human subpneumovirus; and Enterovirus.

[0414] 49. The method according to item 47, wherein the infectious agent is a microorganism selected from the group consisting of: Rickettsia, Chlamydia, Mycobacteria, Clostridia, Corynebacteria, Mycoplasma, Ureaplasma, Legionella, Shigella, Salmonella, pathogenic Escherichia coli species, Bordatella, Neisseria, Treponema, Bacillus, Haemophilus, Moraxella, Vibrio, Staphylococcus spp., Streptococcus spp., Campylobacter spp.), Borrelia spp., Leptospira spp., Erlichia spp., Klebsiella spp., Pseudomonas spp., and Helicobacter pylori spp.

[0415] 50. The method according to item 47, wherein the infectious agent is a microorganism selected from the group consisting of Helicobacter pylori, Chlamydia pneumoniae, Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Streptococcus pneumoniae, Streptococcus viridans, Enterococcus faecalis, Neisseria meningitidis, Neisseria gonorrhoeae, Treponema pallidum, Bacillus anthracis, Salmonella typhi, Vibrio cholerae, cholera), Pasteurella pestis (Yersinia pestis), Pseudomonas aeruginosa, Campylobacter jejuni, Clostridium dificile, Clostridium botulinum, Mycobacterium tuberculosis, Borrelia burgdorferi, Haemophilus ducreyi, Corynebacterium diphtheria, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenza, and enterotoxic Escherichia coli.

[0416] 51. The method of claim 45, wherein the condition comprises an autoimmune disease selected from the group consisting of diabetes, psoriasis of the skin, hyperthyroidism, autoimmune adrenal insufficiency, hemolytic anemia, rheumatic myocarditis, systemic lupus erythematosus, cutaneous lupus erythematosus, psoriatic arthritis, Sjögren's syndrome polymyositis, myositis, multiple sclerosis, Crohn's disease, ulcerative colitis, lupus, inflammatory bowel syndrome, dermatomyositis, dermatitis, atopic dermatitis, scleroderma, polymyositis, psoriasis, alopecia areata, rheumatoid arthritis, graft versus host disease, interferon disease, irritable bowel syndrome, and a combination of two or more of these.

[0417] 52. A method according to item 45, wherein the disease comprises an inflammatory condition, and the inflammatory condition comprises acute kidney injury.

[0418] 53. The method of claim 45, wherein the condition comprises a respiratory disorder comprising asthma, pulmonary fibrosis, or a combination of asthma and pulmonary fibrosis.

[0419] 54. The method according to item 45, wherein the disorder comprises a skin disorder selected from scleroderma, psoriasis, dermatomyositis, atopic dermatitis, and a combination of two or more of these diseases.

[0420] 55. The method according to item 45, wherein the disease comprises a cancer selected from the group consisting of breast cancer, prostate cancer, lymphoma, skin cancer, pancreatic cancer, colon cancer, melanoma, malignant melanoma, ovarian cancer, brain cancer, primary brain cancer, head and neck cancer, glioma, glioblastoma, liver cancer, bladder cancer, non-small cell lung cancer, head and neck cancer, breast cancer, ovarian cancer, lung cancer, small cell lung cancer, Wilms' tumor, cervical cancer, testicular cancer, bladder cancer, pancreatic cancer, gastric cancer, colon cancer, prostate cancer, genitourinary cancer, thyroid cancer, esophageal cancer, myeloma, multiple myeloma, adrenal cancer, renal cell carcinoma, endometrial cancer, adrenocortical carcinoma, malignant pancreatic insulinoma, malignant carcinoid, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, B-cell acute lymphoblastic leukemia, leukemia, blastic plasmacytoid dendritic cell neoplasm (“BPDCN” or “pDC leukemia”), chronic lymphocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia (“AML”), chronic myeloid leukemia, acute myeloid leukemia, senescence-associated hairy cell leukemia, chronic myelomonocytic leukemia (“CMML”), myelodysplastic syndrome, chronic myelofibrosis, multiple myeloma, neuroblastoma, rhabdomyosarcoma, Kaposi’s sarcoma, Waldenstrom’s macroglobulinemia, diffuse large B-cell lymphoma (DLBCL) (L265P), polycythemia vera, essential thrombocythemia, Hodgkin’s disease, non-Hodgkin’s lymphoma, soft tissue sarcoma, osteosarcoma, essential macroglobulinemia, retinoblastoma, and combinations of two or more thereof.

[0421] 56. The method according to items 44 to 55, wherein the epitope comprises the amino acid sequence given by SEQ ID NO: 11, and the antibody or fragment thereof binds to the epitope with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binding epitope.

[0422] 57. The method according to items 44 to 56, wherein the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

[0423] 58. The method according to items 44 to 57, wherein the antibody or fragment thereof is a humanized antibody or fragment thereof.

[0424] 59. The method according to items 44 to 58, wherein:

[0425] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0426] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0427] 60. The method according to items 44 to 58, wherein:

[0428] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0429] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0430] 61. The method according to items 44 to 58, wherein:

[0431] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0432] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0433] 62. The method according to items 44 to 58, wherein:

[0434] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0435] The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:10.

[0436] 63. The method according to items 44 to 58, wherein:

[0437] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0438] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0439] 64. The method according to items 44 to 58, wherein:

[0440] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0441] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0442] 65. The method according to items 44 to 58, wherein:

[0443] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0444] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0445] 66. The method according to items 44 to 58, wherein:

[0446] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0447] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0448] 67. The method according to items 44 to 58, wherein:

[0449] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0450] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0451] 68. The method according to items 44 to 58, wherein:

[0452] The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0453] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0454] 69. A method according to item 44 to 59 or 63, wherein:

[0455] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0456] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0457] 70. A method according to item 44 to 58, 60 or 63, wherein:

[0458] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0459] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0460] 71. A method according to item 44 to 58, 61 or 63, wherein:

[0461] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0462] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0463] 72. A method according to item 44 to 58, 62 or 63, wherein:

[0464] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 4, or a sequence at least about 90% identical to SEQ ID NO: 4; and

[0465] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:5.

[0466] 73. A method according to item 44 to 59 or 64, wherein:

[0467] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0468] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0469] 74. A method according to item 44 to 59 or 65, wherein:

[0470] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0471] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0472] 75. A method according to item 44 to 59 or 66, wherein:

[0473] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0474] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0475] 76. A method according to item 44 to 59 or 67, wherein:

[0476] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0477] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0478] 77. A method according to item 44 to 59 or 68, wherein:

[0479] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and

[0480] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0481] 78. A method according to item 44 to 58, 60 or 64, wherein:

[0482] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0483] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0484] 79. A method according to item 44 to 58, 61 or 64, wherein:

[0485] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0486] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0487] 80. A method according to item 44 to 58, 62 or 64, wherein:

[0488] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 4, or a sequence at least about 90% identical to SEQ ID NO: 4; and

[0489] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:6.

[0490] 81. A method according to item 44 to 58, 60 or 65, wherein:

[0491] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0492] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0493] 82. A method according to item 44 to 58, 61 or 65, wherein:

[0494] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0495] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0496] 83. A method according to item 44 to 58, 62 or 65, wherein:

[0497] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0498] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:7.

[0499] 84. A method according to item 44 to 58, 60 or 66, wherein:

[0500] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0501] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0502] 85. A method according to item 44 to 58, 61 or 66, wherein:

[0503] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0504] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0505] 86. A method according to item 44 to 58, 62 or 66, wherein:

[0506] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0507] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:8.

[0508] 87. A method according to item 44 to 58, 60 or 67, wherein:

[0509] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0510] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0511] 88. A method according to item 44 to 58, 61 or 67, wherein:

[0512] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0513] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0514] 89. A method according to item 44 to 58, 62 or 67, wherein:

[0515] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 4, or a sequence at least about 90% identical to SEQ ID NO: 4; and

[0516] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:9.

[0517] 90. A method according to item 44 to 58, 60 or 68, wherein:

[0518] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and

[0519] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0520] 91. A method according to item 44 to 58, 61 or 68, wherein:

[0521] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and

[0522] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0523] 92. A method according to item 44 to 58, 62 or 68, wherein:

[0524] The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and

[0525] The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

[0526] 93. A kit for depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDCs) in a patient in need thereof, the kit comprising:

[0527] Instructions; and

[0528] An antibody or a fragment thereof, comprising:

[0529] a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0530] A light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0531] 94. A pharmaceutical composition comprising:

[0532] at least one excipient; and

[0533] An antibody or a fragment thereof, comprising:

[0534] a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and

[0535] A light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO: 10.

[0536] 95. Use of the antibody or fragment thereof according to item 1 for treatment.

[0537] 96. The antibody or fragment thereof according to item 1, for use in depleting, inhibiting, or depleting and inhibiting plasmacytoid dendritic cells (pDC).

[0538] 97. Use of the antibody or fragment thereof according to item 1 in the preparation of a medicament.

[0539] Example

[0540] The following describes embodiments related to the present disclosure. In most cases, alternative techniques can be used. These embodiments are for illustrative purposes only and do not limit or restrict the scope of the invention described in the claims.

[0541] In order to determine the role of tumor-associated / TA-pDCs that cause immunosuppression in the human breast cancer tumor microenvironment ("BC-TME") and to elucidate their immune mechanisms, as well as to develop novel therapies (combining pDC-depleting antibodies with other immunotherapies), a fully humanized antibody (NPB-303) was developed that specifically binds to human BDCA2 and inhibits / depletes human pDCs in vivo and in vitro. In a humanized mouse model with human immune and human BC tumor cells ("BC-HIT mice"), it has been demonstrated that TA-pDCs in the BC-TME play a role in the immunosuppressive TME, and that depleting TA-pDCs in the BC-TME reverses immunosuppression, thereby controlling the growth of human BC in vivo. These findings can serve as the basis for clinical studies to test whether TA-pDC depletion will exhibit enhanced anti-tumor activity in BC patients with or without other immuno-oncology ("IO") drugs. The mechanisms of TA-pDCs in programming BC-TME can be studied, and novel immunotherapies in human BC-HIT mice can be simulated.

[0542] Example 1

[0543] Humanization of mouse monoclonal antibody 12b

[0544] The cDNA sequences of the heavy chain variable region ("VH") and light chain variable region ("VL") of mouse monoclonal antibody 12b were determined. These sequences were used to design optimized VH and VL region expression constructs, in which codons rarely used in mammals were replaced by frequently used alternative codons. These gene constructs were synthesized and cloned into human IgG1 antibody expression vectors. Accordingly, the VH and VL region genes were cloned upstream of the heavy chain constant region (CH1, hinge, CH2 and CH3) or the light chain constant region (VL / κ). The resulting chimeric antibody was named NPB-301.

[0545] A three-dimensional molecular model of the mouse 12b variable region was constructed using a proprietary algorithm from JN Biosciences. This was used to identify the framework amino acid residues involved in the formation of the CDR structure. The mouse VH and VL domains from 12b were searched in the database to identify highly homologous human VH and VL domains and selected as recipients for CDR transplantation. Finally, a gene construct was designed in which the 12b CDRs were transplanted onto the human acceptor framework (i.e., replacing the original human CDRs) and the human framework residues corresponding to each structural position in 12b were backmutated to the mouse sequence to ensure functional CDR orientation.

[0546] The genes encoding the humanized VH and VL domains were cloned into an IgG1 expression vector as described above for chimeric antibodies. The resulting humanized antibody variants were named NPB-30X, with different humanized variants being represented by different suffix numbers (i.e., NPB-302, NPB-303, etc.).

[0547] Figure 1 A schematic diagram of an expression vector for producing chimeric and humanized anti-BDCA2 antibodies is provided. The plasmid contains a heavy chain transcription unit initiated with a human CMV promoter (CMV-P) to initiate transcription of the antibody heavy chain gene. The CMV promoter is followed by VH, CH1, hinge, CH2, and CH3 exons. After the heavy chain gene sequence, the light chain transcription unit is initiated with a CMV promoter, followed by VL exons and CL (human kappa chain constant region) exons. The light chain gene is then followed by the SV40 early promoter (SV40-P), the puromycin N-acetyltransferase gene (puro) for puromycin resistance, and a fragment containing the SV40 polyadenylation site (SV40-A). Finally, the plasmid contains a portion of the plasmid pUC19, including a bacterial origin of replication (pUC ori) and a β-lactamase gene (βlactamase). The arrow indicates the direction of transcription. Figure 1 Relevant restriction enzyme sites are also shown.

[0548] In order to obtain stable antibody-producing cell lines, Figure 1The expression vector was stably transfected into CHO-K1 cells (ATCC). Briefly, CHO-K1 cells were cultured in SFM4CHO medium (HyClone) at 37°C and 7.5% CO2. For transfection, 20 μg of the expression vector was linearized using the FspI restriction enzyme and transferred to approximately 10 7 The transfected cells were diluted in SFM4 CHO medium and aliquoted in several 96-well plates. After 48 hours, 10 μg / ml puromycin was added to select for stable integrants. The cells were maintained under puromycin selection for 10 days.

[0549] Antibody expression was measured by ELISA. In brief, the plate was coated with a human IgG Fcγ chain-specific polyclonal antibody (Sigma-Aldrich). The wells were then blocked with PBS containing 0.05% Tween20 and 2% milk. Each well was washed and 100 μL of supernatant was taken out from each stable cell line. The supernatant was added and incubated for 60 minutes. The supernatant was removed by washing, and a HRP-conjugated goat anti-human κ chain polyclonal antibody (Southern Biotech) was added and incubated for 30 minutes. After the last wash, the ELISA signal was generated by adding ABTS substrate and quenched with 2% oxalic acid.

[0550] The detection signal was analyzed at 405 nM using a plate reader. All antibodies were diluted in PBS containing 0.1% skim milk and 0.025% Tween 20. Between each incubation and signal development step, washing was performed using PBS-T (PBS containing 0.05% Tween 20).

[0551] Stable transfectants were cultured in SFM4CHO culture medium using roller bottles until cell viability was less than 50%. Supernatants were collected and clarified by centrifugation and filtration. Samples were loaded onto protein A Sepharose columns (HiTrap MabSelectXtra, GE Healthcare). The columns were washed with PBS and the capture antibodies were eluted with 0.1M glycine-HCl (pH 3). The eluted antibodies were neutralized with 1M Tris-HCl (pH 8), and the buffer was exchanged into PBS by dialysis. Antibody concentration (1mg / ml=1.4OD) was determined by measuring the absorbance at 280nm. Purified NPB-301, NPB-302 and NPB-303 were characterized by SDS-PAGE under reducing conditions according to standard procedures. Figure 2SDS-PAGE analysis of NPB-303 is provided. Figure 3 provides protein analysis of NPB-303 on a Superose 6 column using an AKTA FPLC system. Panel A provides the results of a gel filtration standard generated by BioRad. Panel B provides the results of NPB-303. The preparation has greater than 98% antibody monomer, with a molecular weight of 155 kD.

[0552] Recombinant human and rhesus BDCA2 proteins were generated by Mass Biologics and their binding affinity to anti-BDCA2 antibodies was measured by surface plasmon resonance (SPR) experiments. The extracellular domain of human BDCA2 (UniProt Q8WTT0, residues 45–213) or rhesus BDCA2 (UniProt A0A1D5QFY5, residues 45–212) was cloned into the pYW7 vector or pcDNA3.4 from Mass Biologics, respectively. Proteins were expressed using the ExpiCHO Expression System (Thermo Fisher Scientific). Affinity chromatography purification was performed using a Ni Sepharose Excel column (GE Life Sciences), followed by a standard size exclusion chromatography step to remove aggregates and degradation products.

[0553] The binding kinetics of BDCA2 antibodies were measured using a Biacore surface plasmon resonance (SPR) instrument (GE Life Sciences). Briefly, anti-BDCA2 antibodies were captured on a protein A sensor chip (GE Life Sciences). Recombinant BDCA2 protein was then flowed through a range of concentrations of capture antibody at a rate of 50 μL / min. The data were analyzed using the BIAevaluation software package (GE Life Sciences) to determine the kinetic constants.

[0554] NPB-303 was selected for further study. The heavy chain sequence and light chain sequence of NPB-303 are given by SEQ ID NO: 13 and SEQ ID NO: 14, respectively.

[0555] Figure 4 Biophysical binding of 15b, 12b and NPB-303 to BDCA2 antigen from monkey (panel A) or human (panel B) is shown. The binding data are also summarized in Table 1. In Table 1, monoclonal antibody (mAb) 15b is a mouse monoclonal antibody specific for human BDCA2, mAb 12b is a mouse monoclonal antibody that was humanized to generate NPB-303, which binds to both human and monkey BDCA2, and NPB-303 is a humanized monoclonal antibody.

[0556] Table 1.

[0557]

[0558] Figure 5 The yellow shaded area is the NPB-303 epitope, and the orange shaded area is the BDCA2 lectin (ligand) binding residue. Figure 6 This is a structural model of the variable (binding) domain of NPB-303. In the figure, green represents the VH framework region; yellow represents the VH CDR; blue represents the VL framework region; brown represents the VL CDR; and red represents the aspartate isomerization motif.

[0559] Example 2

[0560] In vitro and in vivo binding characterization of the NPB-303 antibody

[0561] An ELISA assay for NPB-303 binding was developed in-house. Briefly, Nunc MaxiSorp 96-well plates were coated with BDCA-2 antigen (Biorbyt CLEC4C orb358925) overnight at 4°C. After washing, the plates were blocked with blocking buffer (1x PBS containing 1% BSA). Diluted standard NPB-303 antibodies and other test batches were added to the plates. After incubation for 1 hour at room temperature, the plates were washed and secondary antibodies (HRP-conjugated goat anti-human IgG Fc antibody; ThermoFisherA18829) were added and incubated for 1 hour at room temperature. Signals were generated by adding TMB substrate reagent (ThermoFisher 34021). Signals were read at 450nm and 590nm on a plate reader.

[0562] Using LIGHTNING-LINK TM Antibody labeling kit (Novus Biologicals, Cat#765-0005) was used to conjugate NPB-303 antibody and human IgG control antibody with APCCy7 label according to the instructions in the kit. TM One million fresh or frozen human PBMCs were stained with dye (Aqua Yellow) and supplemented with NPB-303-APCCy7 or human IgG-APCCy7, as well as human CD45-BV786, CD3 / CD14 / CD19-FITC, CD4-PacBlue, CD123-PE, and BDCA2-APC. Cells were acquired on a BD FACSCelesta and analyzed using the FlowJo program.

[0563] Umbilical cord blood was obtained from the National Disease Research Interchange (NDRI, Philadelphia, Pennsylvania), and its use was exempted by the New England Institutional Review Board (IRB). TM Human hematopoietic stem cells (HSCs) were enriched with human CD3 depletion cocktail (Stem Cell Technologies). Cells were stained with Viability eF780 (BioLegend), FITC CD3 (BioLegend), PE CD34 (BioLegend), APC CD133 / 2 (Miltenyi) and analyzed on a BD Accuri flow cytometer. Viable cells were counted using ViCell (Beckman Coulter). All cells were isolated under sterile conditions. To construct humanized mice, 1×10 5 Up to 5×10 5 -CD34+CD133+ umbilical cord blood cells were injected intravenously with 1 μg / mL OKT3 mAb added into 4-6 week old pre-irradiated mice. Ten to twelve weeks after implantation, mice were analyzed for reconstitution by flow cytometric analysis of peripheral blood. Mice with 20% or more human leukocyte reconstitution were used in subsequent experiments.

[0564] All experiments involving animals were approved by the Institutional Animal Care and Use Committee (IACUC) of NPB. Female immunodeficient mice, including NSG mice (NOD.Cg-Prkdc scid Iq tm1Wjl / SzJ) and NBSGW (NOD.Cg-Kit from Jackson Laboratory W-41J Tyr + Prkdc scid Iq tm1Wjl / ThomJ) and NP1 (NOD-Rag1- / -IL2RgammaC-null) mice from NPB-USA were used to construct humanized mice. NSG mice and NP1 mice were irradiated at least 4 hours before reconstruction (using Xrad IR225 X-ray irradiator, 180 rads for NSG mice and 400 rads for NP1 mice). Some experiments were performed using humanized NSG mice purchased from Jackson Laboratory (IHCM-humanized CD34 population, implantation rate> 60%).

[0565] Humanized mice were injected with NPB-303 (10 mg / kg IV) on days 0 and 2. Mice were sacrificed on day 3. Live / Dead TMCells from blood were stained with Fixable Yellow (Catalog No. L34959, Thermo Fisher), human CD45-BV786, mouse CD45.1-APCCy7, human CD3 / CD14 / CD19-FITC, human CD4-PacBlue, CD123-PE, and BDCA2-APC (all from BioLegend) and analyzed using the FlowJo program.

[0566] Human PBMCs were incubated with NPB-303 antibody or human IgG1 isotype control mAb (10 μg / mL) at 37°C for 30 minutes. PBMCs were then stimulated with CpG-ODN-2216 (Invivogen, catalog number tlrl-2216, 2 μg / mL) for 16 hours. Cells were treated with GolgiPlug for 3 hours before flow cytometry analysis. After culture, human PBMCs were stained with surface markers including Live-Dead Yellow Dye, human CD45-BV786, CD3-FITC, CD14-FITC, CD19-FITC, CD123-PECy7, BDCA2-APC, CD4-Pacific Blue, and then intracellular staining was performed for IFNα-PE or IgG1-PE isotype controls using fixation / permeabilization solution (BD Cytofix / Cytoperm, catalog number 54714). The stained cells were analyzed using a BD Celesta flow cytometer and the data were analyzed using FlowJo software.

[0567] 1 x 10 cells / mL were plated in complete RPMI medium (10% FBS, 1 x non-essential amino acids, 1 x Pen-Strp, 1 mM sodium pyruvate, 10 mM HEPES, 50 μM 2-mercaptoethanol, and 2 mM L-glutamine) in 96-well round-bottom plates. 6 / well of human PBMC and treated with 10, 3.33, 1.11, 0.37, 0.124, 0.04, 0.014, 0.005, 0.0015 and 0.0005 μg / mL of NPB-303 antibody or 10 μg / mL of isotype control antibody for 1 hour. All conditions were established in duplicate. The cells were then stimulated with CpG-ODN-2216 at a final concentration of 10 mg / mL and cultured for 16 hours at 37 ° C and 5% CO2. The supernatant was collected and IFNα was evaluated using the PBL Human IFNAlpha Multi-Subtype ELISA Kit (Serum, Plasma, TCM). IC was calculated using the GraphPad Prism program 50 .

[0568] Human PBMCs were incubated with NBP-303 antibody or human IgG1 isotype control mAb (10 μg / mL) at 37°C for 30 minutes. PBMCs were then stimulated with CpG-ODN-2216 (Invivogen, catalog number tlrl-2216, 2 μg / mL) for 16 hours. Cells were treated with GolgiPlug for 3 hours before flow cytometry analysis. After culture, human PBMCs were stained with surface markers including Live-Dead Yellow Dye, human CD45-BV786, CD3-FITC, CD14-FITC, CD19-FITC, CD123-PECy7, BDCA2-APC, CD4-Pacific Blue, and then intracellular staining was performed for IFNα-PE or IgG1-PE isotype controls using fixation / permeabilization solution (BD Cytofix / Cytoperm, catalog number 54714). The stained cells were analyzed using a BD Celesta flow cytometer and the data were analyzed using FlowJo software.

[0569] NPB-303 binds to BDCA2 on human pDCs (in vitro) and depletes / inhibits pDCs in vivo. To elucidate the role of the chronic pDC / IFN-I axis in suppressing antitumor immunity in the tumor microenvironment, a novel humanized antibody was developed to target human pDCs for the treatment of human inflammatory diseases associated with TA-pDCs. A murine monoclonal antibody that specifically binds to human BDCA2 on pDCs was engineered by replacing the non-antigen binding regions of the Fc domain and Fab domain with human antibody sequences (see Figure 1 NPB-303 antibody (huIgG1) showed a highly stable interaction with BDCA2 protein ( Figure 7 )( Figure 8 ,K d About 1nM,k on (1 / ms) Approx. 2.1x10 5 , k off (1 / s)about 3.3x10 -4 NPB-303 specifically binds to human pDC in vitro ( Fig. 9 The inhibition of BDCA2-mediated pDC activation by NPB-303 was examined in vitro. NPB303 effectively inhibited CpG-activated pDCs, as detected by intracellular IFNα ( Fig.10 ) or by detection of IFNα in the supernatant ( Fig.11 ). Similar studies were performed in multiple human donor PBMC samples, and IC 50 Estimated to be about 3ng / ml ( Fig.12 and Fig.13), significantly superior to the recently reported Biogen BDCA2Mab.

[0570] Example 3

[0571] Pharmacokinetic and pharmacodynamic characterization of NPB-303 antibody

[0572] All animals used were NOD Rag Gamma (NRG) mice purchased from Jackson Laboratories (Catalog No. 007799). All animals were male and 5–6 weeks old. Animals were injected with NPB-303 at a concentration of 10 mg / mL. The dose was 10 mL / kg, resulting in an antibody dose of 10 mg / kg. Mice were dosed once by intravenous (IV) (tail vein) or subcutaneous (SC) (flank) injection at the beginning of the experiment. Blood was collected by cheek bleed at different time points after NPB-303 injection. Groups of three animals were treated for each time point. Sample volume was limited to 50 μL per blood draw during the survival period and to a total of approximately 200 μL (10% of blood volume) during the two-week study period. Prior to the last sample collection from a given animal, mice were euthanized by CO2 asphyxiation and terminal blood was collected by cardiac puncture. Blood was collected in K3EDTA tubes (Greiner Bio-One) and processed into plasma.

[0573] NPB-303 was detected in plasma samples using an IgG (total) human non-coated ELISA kit (ThermoFisher catalog number: 88-50550-22) according to the manufacturer's instructions. A pilot optimization was performed using 16 samples (n=1) at 5 different dilutions: 1:50, 1:2000, 1:500, 1:5000, and 1:10,000. The samples tested also included a standard curve constructed by preparing a 100 ng / mL solution of purified NPB-303, which was then used to generate a 2-fold serial dilution (a total of 7 concentrations ranging from approximately 1.6–100 ng / mL). The NPB-303 concentration in the plasma samples was estimated by comparison with this curve. The data were used to calculate the half-life (T 1 / 2 ), maximum plasma concentration (C max ), time to reach maximum plasma concentration (T max ), mean residence time (MRT), area under the plasma concentration curve (AUC), apparent systemic clearance (CL) and apparent volume of distribution at the terminal stage (VZ). All data analyses were performed using WinNonlin 7.0 software package (Certara).

[0574] NPB-303 was further characterized to quantify the presence of endotoxin.

[0575] Humanized female mice were injected intravenously with 10 mg / kg of NPB-303. Mice were sacrificed at 0, 1, 6, 24, 48, 72, and 168 hours after injection. Mouse splenocytes were stained with Live / Dead Dye, mouse CD45.1 APCCy7, and human antibodies, including CD3 FITC, CD14FITC, CD19 FITC, CD123 PE, BDCA2 APC, CD4 Pacific Blue, CD45 BV786 (all from BioLegend). Cells were analyzed on a BD Celesta instrument and data were analyzed using FlowJo software.

[0576] The pharmacokinetics of NPB-303 in NSG mice were studied by intravenous or subcutaneous injection. Fig.14 and Table 2). In Table 2, T 1 / 2 is the terminal half-life of the analyte in plasma, T max is the time from drug administration to the measurement of maximum concentration, C max is the maximum measured concentration of the analyte in plasma, MRT is the mean residence time, and AUC 0-∞ is the area under the concentration-time curve of the analyte in plasma over the time interval extrapolated from 0 to infinity, CL is the clearance, V Z is the apparent volume of distribution at the end stage. Fig.14 In the figure, the cross on the "Time Point (hr)" axis represents the plasma concentration of NPB-303 detected in the sample collected 30 minutes before antibody injection. The estimated half-life of NPB-303 is 6.2 days for intravenous injection and 4.2 days for subcutaneous injection, respectively. For intravenous and subcutaneous routes, T max 1 hour and 36 hours respectively, C max 109 μg / ml and 80 μg / ml, respectively. No adverse effects were detected. Therefore, the calculated values ​​of these parameters provided in Table 2 are highly dependent on the sampling protocol.

[0577] Table 2.

[0578]

[0579] When tested in mice transplanted with human CD34+ HSC / HSPC, NPB-303 potently and specifically depleted human pDCs in humanized mice in vivo ( Fig.15 , Fig.16 , Fig.17 and Fig.18 ). 10 days after mAb treatment, BDCA2 surface protein was efficiently internalized onto the remaining pDCs ( Fig.19 Human pDCs in lymphoid organs are effectively depleted and / or inhibited by BDCA2 binding ( Fig.19 and Fig. 20 ). Therefore, humanized NPB-303 mAb can specifically bind to BDCA2 on human pDCs and effectively inhibit and deplete pDCs in vivo and in vitro. However, NPB-303 did not deplete other cell types in the blood (Lin-CD4-, CD4+CD123- and CD4-CD123-), such as Fig.21 shown.

[0580] Fig. 22 Endotoxin testing using a kinetic turbidimetric and kinetic chromogenic test (ENDOSAFE) is provided. The endotoxin level of NPB-303 (1:1000 dilution) is less than 0.005 EU / mL, and the reaction time is approximately 2520 seconds.

[0581] Example 4

[0582] Antitumor activity of NPB-303 antibody

[0583] Two different human triple negative breast cancer (TNBC) cell lines were purchased from the American Type Culture Collection (ATCC): MDA-MB-231 (ATCC HTB-26) and MDA-MB-468 (ATCC HTB-132). Cells were cultured in Leibovitz's L-15 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (GE Healthcare Life Sciences, HyClone Laboratories) and 1% penicillin-streptomycin (Thermo Fisher Scientific) at 37°C in 5% CO2.

[0584] At least 12 weeks after human HSC transplantation, 5x10 6 TNBC MDA-MB-231 or MDA-MB-468 cells were mixed with Matrigel (Corning Life Sciences) and injected into the thoracic mammary fat pad of female humanized mice. Tumor volume was monitored weekly using a digital caliper.

[0585] Once tumor size in humanized mice bearing tumors (MDA-MB-231) reached approximately 100 mm 3 , about 300mm 3 or about 600mm 3 Mice were sacrificed and the infiltration of human pDCs in tumor tissue was analyzed using a previously described flow cytometry pDC panel. In a separate experiment, once tumors reached approximately 300 mm in size,3 , then blood, spleen, tumor, lung and liver were separated and analyzed for human pDC and T cell infiltration. For T cell flow cytometry analysis, cells were stained with Live / Dead Dye, mouse CD45.1 APCCy7 and human antibodies, including CD3 PECy7, CD4 PE, CD8 BV515, PD1 BV421 and Tim3 APC (all from BioLegend). Cells were analyzed on a BD FACSCelesta instrument and data were analyzed using FlowJo software.

[0586] Once the tumor reaches approximately 80–100 mm 3 Humanized NSG mice carrying TNBC (MDA-MB-231) were randomly divided into 4 treatment groups. Mice were treated with NPB-303 or isotype control antibody (30 mg / kg) intravenously twice a week at 3, 10, 30 mg / kg for 4 weeks. Tumor volume, body weight and gross observations were recorded twice a week. At the end of the study, pDC infiltration analysis was performed on spleens and tumors as described previously.

[0587] For the development of the second TNBC (MDA-MB-468) model, HIT-TNBC mice were randomized and intravenously injected with NPB-303 (at doses of 1 and 10 mg / kg) and isotype control antibody (10 mg / kg) twice a week for three weeks. Tumor volume, body weight, and gross observations were recorded twice a week.

[0588] For the combination study, humanized NBSGW (NOD.Cg-Kit) carrying TNBC (MDA-MB-231) was used. W-41J Tyr + Prkdc scid Iq tm1Wjl / ThomJ) mice. Once the tumor volume reached approximately 200 mm3, the mice were randomized into four treatment groups: isotype control 30 mg / kg, intravenously injected twice a week for 3 weeks; alone, 5 mg / kg, intraperitoneally, once daily for 5 days only; NPB-303, 30 mg / kg, intravenously, twice weekly for 3 weeks; and The combination of 5 mg / kg and NPB-303 (30 mg / kg) was treated at the same frequency as the single treatment groups. Tumor volume was recorded twice a week. At the end of the study, mice were sacrificed and cells in the spleen, tumors, and blood were analyzed. Liver tissue was also isolated for histological analysis.

[0589] Liver tissues were removed from mice in the combined studies and immediately fixed in 4% paraformaldehyde for up to 48 hours. The tissues were then moved to 70% ethanol until embedded. Each tissue was sectioned every 100 μm [micrometers], typically 10 sections per organ, and stained with H&E.

[0590] Specimens were evaluated by a board-certified veterinary pathologist (HORUS Scientific & NOVA Preclinical Consultants, LLC) to determine morphological evidence and extent of tumor infiltration into lung and liver tissues. Tumor infiltration was characterized using light microscopy using the following semiquantitative scale: (0) no observable changes; (1) minimal - barely perceptible / inconspicuous features / changes in tissues; (2) slight / subtle features or presence in tissues; (3) moderate / significant features / presence in tissues; and (4) marked / extensive features / presence in tissues. Tumor metastasis scores for all specimens were generated in Microsoft EXCEL TM The data were analyzed and the p-values ​​between groups were calculated using t-test.

[0591] Given the functional human immunity developed in NSG / NRG-hu HSC mice, these mice partially reject subcutaneously co-implanted human tumors. NSG-hu HSC mice support the growth of non-HLA-matched tumors and can enhance human immune cells to control tumor growth. Described herein are humanized mice with human immune cells and tumors ("HIT mice").

[0592] In addition, immuno-oncology checkpoint inhibitors anti-PD1 mAb NPB-303 showed similar efficacy in tumor-bearing humice A comparable reduction in tumor volume ( Fig.23 ), and tumor-bearing humice appeared to tolerate NPB-303 treatment well ( Fig.24 ). However, spleen and tumor pDCs were depleted by NPB-303, but not by Consumption( Fig.25 and Fig.26 ). Similarly, spleen and tumor BDCA-2 were downregulated by NPB-303, but not Downward adjustment Fig. 27 and Fig.28 ).

[0593] like Fig.29 As shown, using HIT mice with human triple-negative breast cancer cells ("TNBC-HIT"), the results showed that transplanted tumors were infiltrated with human CD45 leukocytes, including CD123+CD4+CD303+pDCs ( Fig.30 and Fig.31 In addition, markers of immune exhaustion (PD1 and TIM3, Fig.32 , Figure 339 and Fig.34 ) of human T cells.

[0594] NPB-303 was examined in the TNBC-HIT model for the treatment of human TNBC ( Fig.35 ). Carry about 100mm 3 MDA-MB-231TNBC-bearing HIT mice were treated with NPB-303 or isotype control mAb (30 mg / kg) at doses of 3 mg / kg, 10 mg / kg, and 30 mg / kg, and tumor growth was monitored for 31 days. Tumor volumes in all NPB-303-treated groups were significantly smaller than those in the isotype control mAb-treated group, and a dose-dependent response was observed ( Fig.35 No obvious adverse effects were observed with mAb ( Fig.36 As expected, human pDCs were expressed in tumors of NPB-303-treated mice ( Fig.37 ) and spleen ( Fig.38 The remaining pDCs also lost BDCA2 / CD303 expression, indicating that NPB-303 binds to and inhibits all human pDCs.

[0595] To confirm the therapeutic activity of NPB-303 against human TNBC, we tested its activity in TNBC cells transplanted with another human TNBC cell line (MDA-MB-468, Fig.39 Similar tumor reduction activity was observed in MB-468-HIT mice ( Fig.40 ) and had no significant adverse effects on HIT mice ( Fig.41 ).

[0596] Current treatments for TNBC patients include First-line chemotherapy followed by immune checkpoint inhibitors such as anti-PD1 mAb Therefore, we tested the Combination activity with NPB-303 after treatment ( Fig.42 ). When tracking tumor size after treatment, or NPB-303 inhibited tumor growth to similar sizes (about 50%, Fig.43 Importantly, the Abx / NPB-303 combination further reduced tumor size by approximately 90% ( Fig.43 ) and had no significant adverse effects on HIT mice ( Fig.44 ). Compared with using ISO or Compared with the treatment groups, the use of NPB-303 or NPB-303 and Combination therapy can significantly reverse tumor-infiltrating CD8 T cells by downregulating PD1 and Tim3 expression ( Fig.45 and Fig.46 ).

[0597] Treatment may lead to tumor metastasis. Therefore, metastatic tumors ( Fig.43 ). The treated mice showed significant tumor metastasis to the liver, while the NPB-303-treated mice had no or less tumor metastasis. and NPB-303 showed liver ( Fig.47 , Fig.48 and Fig.49 ) and lungs ( Fig.50 and Fig.51 ) significantly reduced tumor metastasis. These data suggest that NPB303-induced tumor suppression is independent of enhanced tumor metastasis and that NPB-303 is The combination synergistically inhibits tumor growth and reduces tumor metastasis.

[0598] NPB-303 also exhibits thermal stability characteristics similar to other clinical-stage and marketed monoclonal antibodies. Fig.52 As shown, the thermal denaturation (T m ) and aggregation occurs (T agg ) is comparable to therapeutic antibodies that have at least progressed to clinical trials. In addition, NPB-303 binding activity was stable under various heat stress conditions. Fig.53 The binding activity of NPB-303 under different conditions is shown. There is no difference between freshly thawed NPB-303, NPB-303 incubated at 4°C for 2 weeks, and NPB-303 subjected to stress incubation at 42°C for 2 weeks.

[0599] pDC / IFN-I in the tumor microenvironment is associated with immunosuppression. NPB-303 inhibited TA-pDC in a dose-dependent manner in TNBC-HIT mice, which was confirmed by using TNBC-HIT mice with two human TNBC cell lines (MB231 and MB468). NPB-303 was examined in combination with first-line chemotherapy drugs. The effect of combined use. Either PD-1 or NPB-303 reduced tumor growth, while their combination synergistically inhibited tumors. NPB-303 showed significant activity in preventing tumor metastasis to the liver and lung. ) have shown some efficacy in treating various types of human cancer, but most cancer patients do not respond to immunosuppressive agents due to other mechanisms including persistent inflammation. No response. This example provides a new weapon for the effective treatment of cancer.

[0600] Example 5

[0601] NPB-303 Antibody Developability and Manufacturability Engineering

[0602] With the increasing number of candidate drugs entering clinical trials and development, the evaluation of safety and efficacy has become increasingly important. Compared with other molecular drugs, monoclonal antibodies are more susceptible to various post-translational modifications (PTMs) during production, transportation and storage, which can affect the stability of drug molecules and the structure and function of proteins. Aspartic acid isomerization is a spontaneous non-enzymatic PTM method. After aspartic acid isomerization, a methylene group is inserted into the protein backbone, that is, a methylene group is reduced on the aspartic acid side chain, thereby causing changes in protein structure and protein function. Many studies have revealed that aspartic acid isomerization can affect the effectiveness of candidate drugs. For example, aspartic acid isomerization in the complementarity determining region (CDR) of an antibody can reduce receptor binding efficiency and affect the final drug efficacy.

[0603] In the NPB-303 sequence, DG may be the problematic amino acid causing the isomerization problem. Developability engineering studies were performed by making the following modifications to the antibody sequence: 1) changing DG in the heavy chain CDR3 to DA, EG, and SG without changing the light chain to determine if these changes affect affinity; 2) changing DGDG in the light chain CDR1 to EGEG, DADA, EGDA, DAEG, SGSG (which can be SGDG and DGSG) without changing the heavy chain to determine if these changes affect affinity; 3) combining the best changes in the heavy and light chains in the same antibody molecule. Detailed sequence modifications are shown in Figure 2. Fig.54 As shown, including SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 and SEQ ID NO: 10.

[0604] Antibodies with different heavy and light chain combinations were produced on a small scale (3 mL) using a transient antibody production method (Table 3). After protein purification, the antibodies were analyzed, including titer, A280, SEC-HPLC, and Caliper SDS (reduced and non-reduced). The QC data for all antibodies are summarized in Table 4. All antibodies were checked in a formulation containing 50 mM Na-AC, 33 mM Tris, and the specified amount of antibody at a pH of 5.0.

[0605] Table 3

[0606]

[0607]

[0608] Table 4

[0609]

[0610] The binding kinetics of BDCA2 antibodies were measured using a Biacore surface plasmon resonance (SPR) instrument (GE Life Sciences). Briefly, anti-BDCA2 antibodies were captured on an anti-human Fc chip. Recombinant BDCA2 protein was then flowed through a range of concentrations of capture antibody at a rate of 50 μL / min. The data were analyzed using the BIAevaluation software package (GE LifeSciences) to determine the kinetic constants. Fig.55 The sensorgrams of SPR-based affinity testing of all 24 antibodies are shown in and the biophysical properties of all 24 antibodies are summarized in Table 5 .

[0611] Table 5

[0612]

[0613] Furthermore, the biological activity of these 24 antibodies in inhibiting TLR-9-induced IFNα production was tested in PBMCs from a healthy human donor cultured in vitro. Fig.56 As shown, the potency (IC 50 ) Fig.57 Although the IC 50 There were differences, but all 24 antibodies showed dose-dependent inhibition of TLR-9-induced IFNα production.

[0614] Based on the binding affinity, expression level and potency of all 24 antibodies, the top 3 antibodies (NPB-303_A.D; NPB-303_C.D and NPB-303_C.G) were selected and produced on a large scale (100 mL) using transient transfection. The QC data of the top 3 antibodies are summarized in Table 6. The binding affinity was analyzed using the SPR method, and the sensorgrams of all three antibodies are shown in Table 6. Fig.58 The biophysical activities are summarized in Table 7. The data indicate that the biophysical characteristics of the large-scale production are similar to those of the small-scale production.

[0615] Table 6

[0616]

[0617] Table 7

[0618]

[0619] Large-scale production of NPB-303_A.D; NPB-303_C.D and NPB-303_C.G were also tested for inhibition of TLR-9-induced IFNα production. Dose-dependent IFNα inhibition was observed in two different healthy donors ( Fig.59 ), and obtain Fig.60 The IC shown in 50 value.

[0620] The in vivo human pDC depletion of large-scale produced NPB-303_A.D; NPB-303_C.D and NPB-303_C.G was further tested in humanized mice. Briefly, 20 humanized mice were randomly divided into 5 groups (4 mice per group). Mice were treated with 10 mg / kg of isotype control antibodies NPB-303_WT, NPB-303_A.D, NPB-303_C.D or NPB-303_C.G intravenously. Seventy-two (72) hours later, mice were sacrificed and pDC analysis was performed on blood and spleen.

[0621] Fig.61 Representative flow cytometry plots of blood samples from one mouse in each group are shown in Figure 2. Human pDCs were depleted in all mice treated with NPB-303 antibody compared to isotype controls ( Fig.62 ), and BDCA2 expression was downregulated in all mice treated with NPB-303 antibody ( Fig.63 ). The depletion of pDC in splenocytes was also analyzed by flow cytometry. Fig.64 Flow cytometry of a representative spleen from one mouse in each group is shown in FIG. Fig.65 ) and BDCA2 downregulation ( Fig.66In addition, NPB-303 antibody treatment did not affect the Fig.68 ) and spleen ( Fig.69 ) including Lin-CD4-, CD4+CD123- and CD4-CD123- cells ( Fig.67 ).

[0622] Although embodiments have been disclosed above, the present invention is not limited to the disclosed embodiments. On the contrary, this application is intended to cover any variation, use or adaptation of the present invention using the general principles of the present invention. In addition, this application is intended to cover content that is different from the present disclosure, belongs to known or customary practices in the field to which the present invention belongs, and falls within the scope of the appended claims.

Claims

1. An antibody or a fragment thereof, comprising: a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and A light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, and a sequence that is at least about 90% identical to SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, or SEQ ID NO:

10.

2. The antibody or fragment thereof according to claim 1, wherein the antibody or fragment thereof binds to the molecule with an affinity (K) of 0.1 nanomolar (nM) to 100 nM. d ) binds to the epitope having SEQ ID NO:

11.

3. The antibody or fragment thereof according to claim 1 or claim 2, wherein the antibody or fragment thereof is a monoclonal antibody or fragment thereof.

4. The antibody or fragment thereof according to claim 1 or claim 2, wherein the antibody or fragment thereof is a humanized antibody or fragment thereof.

5. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:

10.

6. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:

10.

7. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:

10.

8. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and a sequence that is at least about 90% identical to SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, or SEQ ID NO:

10.

9. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:

5.

10. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:

6.

11. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:

7.

12. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8.

13. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:

9.

14. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and a sequence that is at least about 90% identical to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:

10.

15. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:

5.

16. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:

5.

17. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:

5.

18. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:5, or a sequence at least about 90% identical to SEQ ID NO:

5.

19. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:

6.

20. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:

7.

21. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8.

22. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:

9.

23. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 1 or a sequence at least about 90% identical to SEQ ID NO: 1; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:

10.

24. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:

6.

25. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:

6.

26. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:6, or a sequence at least about 90% identical to SEQ ID NO:

6.

27. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:

7.

28. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:

7.

29. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:7, or a sequence at least about 90% identical to SEQ ID NO:

7.

30. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8.

31. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8.

32. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:8, or a sequence at least about 90% identical to SEQ ID NO:

8.

33. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:

9.

34. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:

9.

35. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:9, or a sequence at least about 90% identical to SEQ ID NO:

9.

36. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 2 or a sequence at least about 90% identical to SEQ ID NO: 2; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:

10.

37. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO: 3, or a sequence at least about 90% identical to SEQ ID NO: 3; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:

10.

38. The antibody or fragment thereof according to claim 1 or claim 2, wherein: The heavy chain variable region comprises the amino acid sequence given by SEQ ID NO:4, or a sequence at least about 90% identical to SEQ ID NO:4; and The light chain variable region comprises the amino acid sequence given by SEQ ID NO:10, or a sequence at least about 90% identical to SEQ ID NO:10.

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