WO2022257900A1 - 抗masp-2抗体及其用途 - Google Patents

抗masp-2抗体及其用途 Download PDF

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WO2022257900A1
WO2022257900A1 PCT/CN2022/097268 CN2022097268W WO2022257900A1 WO 2022257900 A1 WO2022257900 A1 WO 2022257900A1 CN 2022097268 W CN2022097268 W CN 2022097268W WO 2022257900 A1 WO2022257900 A1 WO 2022257900A1
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seq
amino acid
acid sequence
binding protein
sequence shown
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PCT/CN2022/097268
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English (en)
French (fr)
Inventor
刘小五
曹晓丹
宋剑秋
王宗达
刘培培
张建建
顾春银
邓俗俊
潘忠宗
王学萍
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上海济煜医药科技有限公司
江西济民可信集团有限公司
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Application filed by 上海济煜医药科技有限公司, 江西济民可信集团有限公司 filed Critical 上海济煜医药科技有限公司
Priority to CN202280040887.XA priority Critical patent/CN117460750A/zh
Priority to IL309231A priority patent/IL309231A/en
Priority to AU2022288660A priority patent/AU2022288660A1/en
Priority to KR1020247000299A priority patent/KR20240017071A/ko
Priority to BR112023025596A priority patent/BR112023025596A2/pt
Priority to EP22819498.1A priority patent/EP4353749A1/en
Priority to US18/568,518 priority patent/US20240141069A1/en
Priority to CA3221859A priority patent/CA3221859A1/en
Publication of WO2022257900A1 publication Critical patent/WO2022257900A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
    • C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/565—Complementarity determining region [CDR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/567—Framework region [FR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
    • G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
    • G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
    • G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
    • G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
    • G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
    • G01N2333/96433—Serine endopeptidases (3.4.21)

Definitions

  • This application relates to the field of biomedicine, in particular to an anti-MASP-2 antibody and its application.
  • Immunoglobulin A nephropathy (also known as Berger disease) is a type of mesangial proliferative glomerulonephritis (GN) characterized by diffuse deposition of IgA in the renal mesangium.
  • IgAN immunoglobulin A nephropathy
  • KIDGO mesangial proliferative glomerulonephritis
  • the KIDGO guidelines recommend the use of ACEi/ARB to reduce urinary protein and improve renal function.
  • immunofluorescence studies have shown that local complement C3 activation in glomeruli is associated with poor prognosis, suggesting that IgA nephropathy is related to the activation of the complement system.
  • the complement system consists of three pathways: 1) the classical pathway (CP classical pathway); 2) the alternative pathway (AP alternative pathway); 3) the lectin pathway (LP lectin pathway).
  • CP classical pathway classical pathway
  • AP alternative pathway alternative pathway
  • lectin pathway LP lectin pathway
  • MASP-2 is an effector enzyme of the lectin signaling pathway in the complement system, and is one of the ideal targets to prevent the abnormal activation of complement.
  • the application provides an isolated antigen-binding protein, which has one or more of the following properties: 1) specificity to human MASP-2 protein with a KD value of about 2E-09M or below in the Octet assay Binding; 2) in the Octet assay, specifically binding to the monkey MASP-2 protein with a KD value of about 2E-09M or below; and 3) being able to specifically block the lectin pathway of the human complement system without affecting the classical pathway of complement and bypass pathway.
  • the isolated antigen binding protein comprises HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19.
  • the isolated antigen binding protein comprises HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 17.
  • the isolated antigen binding protein comprises HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 15.
  • the isolated antigen binding protein comprises HCDR1, HCDR2 and HCDR3 of the heavy chain variable region VH shown in SEQ ID NO:67.
  • the isolated antigen binding protein comprises HCDR1, HCDR2 and HCDR3 of the heavy chain variable region VH shown in SEQ ID NO: 13 and SEQ ID NO: 26.
  • the antigen binding protein of described isolation comprises heavy chain variable region VH
  • described VH comprises described HCDR1, HCDR2 and HCDR3, and described HCDR3 comprises the aminoacid sequence shown in SEQ ID NO:19
  • the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 17
  • the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 15.
  • the isolated antigen binding protein comprises H-FR1, the C-terminus of the H-FR1 is directly or indirectly linked to the N-terminus of the HCDR1, and the H-FR1 comprises SEQ ID NO : The amino acid sequence shown in 62.
  • the H-FR1 comprises the amino acid sequence shown in any one of SEQ ID NO:14 and SEQ ID NO:27.
  • said isolated antigen binding protein comprises H-FR2, said H-FR2 is located between said HCDR1 and said HCDR2, and said H-FR2 comprises SEQ ID NO: 63 amino acid sequence.
  • the H-FR2 comprises the amino acid sequence shown in any one of SEQ ID NO:16 and SEQ ID NO:28.
  • the isolated antigen binding protein comprises H-FR3, the H-FR3 is located between the HCDR2 and the HCDR3, and the H-FR3 comprises SEQ ID NO: 64 amino acid sequence.
  • the H-FR3 comprises the amino acid sequence shown in any one of SEQ ID NO:18 and SEQ ID NO:29.
  • the isolated antigen binding protein comprises H-FR4, the N-terminus of the H-FR4 is directly or indirectly linked to the C-terminus of the HCDR3, and the H-FR4 comprises SEQ ID NO : Amino acid sequence shown in 65.
  • the H-FR4 comprises the amino acid sequence shown in any one of SEQ ID NO:20 and SEQ ID NO:30.
  • the isolated antigen-binding protein comprises H-FR1, H-FR2, H-FR3 and H-FR4, and the H-FR1 comprises the amino acid sequence shown in SEQ ID NO: 62; the H-FR2 comprises the amino acid sequence shown in SEQ ID NO: 63; the H-FR3 comprises the amino acid sequence shown in SEQ ID NO: 64; and the H-FR4 comprises the amino acid sequence shown in SEQ ID NO: 65.
  • the H-FR1 comprises the amino acid sequence shown in any one of SEQ ID NO:14 and SEQ ID NO:27;
  • the H-FR2 comprises SEQ ID NO:16 and SEQ ID NO: The amino acid sequence shown in any one of 28;
  • the H-FR3 comprises the amino acid sequence shown in any one of SEQ ID NO:18 and SEQ ID NO:29;
  • the H-FR4 comprises SEQ ID NO:20 and the amino acid sequence shown in any one of SEQ ID NO:30.
  • the H-FR1, H-FR2, H-FR3 and H-FR4 in the isolated antigen binding protein comprise an amino acid sequence selected from any of the following groups:
  • H-FR1 SEQ ID NO:14
  • H-FR2 SEQ ID NO:16
  • H-FR3 SEQ ID NO:18
  • H-FR4 SEQ ID NO:20;
  • H-FR1 SEQ ID NO:27
  • H-FR2 SEQ ID NO:28
  • H-FR3 SEQ ID NO:29
  • H-FR4 SEQ ID NO:30.
  • the isolated antigen binding protein comprises a heavy chain variable region VH comprising the amino acid sequence shown in SEQ ID NO:67.
  • the isolated antigen binding protein comprises a heavy chain variable region VH comprising the amino acid sequence shown in any one of SEQ ID NO:13 and SEQ ID NO:26.
  • the isolated antigen binding protein comprises LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11.
  • the isolated antigen binding protein comprises LCDR2 comprising the amino acid sequence shown in SEQ ID NO:9.
  • the isolated antigen binding protein comprises LCDR1 comprising the amino acid sequence shown in SEQ ID NO:7.
  • the isolated antigen binding protein comprises LCDR1, LCDR2 and LCDR3 of the light chain variable region VL shown in SEQ ID NO:66.
  • the isolated antigen binding protein comprises LCDR1, LCDR2 and LCDR3 of the light chain variable region VL shown in any one of SEQ ID NO:5 and 21.
  • the antigen binding protein of described isolation comprises light chain variable region VL, and described VL comprises described LCDR1, LCDR2 and LCDR3, and described LCDR3 comprises the aminoacid sequence shown in SEQ ID NO:11;
  • the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 9; and the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 7.
  • the isolated antigen binding protein comprises L-FR1, the C-terminus of the L-FR1 is directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 comprises SEQ ID NO : The amino acid sequence shown in 58.
  • the L-FR1 comprises the amino acid sequence shown in any one of SEQ ID NO:6 and SEQ ID NO:22.
  • said isolated antigen binding protein comprises L-FR2, said L-FR2 is located between said LCDR1 and said LCDR2, and said L-FR2 comprises SEQ ID NO:59 amino acid sequence.
  • the L-FR2 comprises the amino acid sequence shown in any one of SEQ ID NO:8 and SEQ ID NO:23.
  • said isolated antigen binding protein comprises L-FR3, said L-FR3 is located between said LCDR2 and said LCDR3, and said L-FR3 comprises SEQ ID NO: 60 amino acid sequence.
  • the L-FR3 comprises the amino acid sequence shown in any one of SEQ ID NO:10 and SEQ ID NO:24.
  • the isolated antigen binding protein comprises L-FR4, the N-terminus of the L-FR4 is directly or indirectly linked to the C-terminus of the LCDR3, and the L-FR4 comprises SEQ ID NO : Amino acid sequence shown in 61.
  • the L-FR4 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:12 and SEQ ID NO:25.
  • the isolated antigen-binding protein comprises L-FR1, L-FR2, L-FR3 and L-FR4, and the L-FR1 comprises the amino acid sequence shown in SEQ ID NO: 58; the L-FR2 comprises the amino acid sequence shown in SEQ ID NO: 59; the L-FR3 comprises the amino acid sequence shown in SEQ ID NO: 60; and the L-FR4 comprises the amino acid sequence shown in SEQ ID NO: 61.
  • the L-FR1 comprises the amino acid sequence shown in any one of SEQ ID NO:6 and SEQ ID NO:22; the L-FR2 comprises SEQ ID NO:8 and SEQ ID NO: The amino acid sequence shown in any one of 23; the L-FR3 comprises the amino acid sequence shown in any one of SEQ ID NO:10 and SEQ ID NO:24; and the L-FR4 comprises SEQ ID NO:12 and the amino acid sequence shown in any one of SEQ ID NO:25.
  • said L-FR1, L-FR2, L-FR3 and L-FR4 in said isolated antigen binding protein comprise an amino acid sequence selected from any of the following groups:
  • L-FR1 SEQ ID NO:6, L-FR2: SEQ ID NO:8, L-FR3: SEQ ID NO:10 and L-FR4: SEQ ID NO:12;
  • L-FR1 SEQ ID NO:22
  • L-FR2 SEQ ID NO:23
  • L-FR3 SEQ ID NO:24
  • L-FR4 SEQ ID NO:25.
  • the VL of the isolated antigen binding protein comprises the amino acid sequence shown in SEQ ID NO:66.
  • the VL comprises the amino acid sequence shown in any one of SEQ ID NO:5 and SEQ ID NO:21.
  • the VH and VL of the isolated antigen binding protein comprise amino acid sequences selected from any of the following groups:
  • VH SEQ ID NO: 13 and VL: SEQ ID NO: 5;
  • VH SEQ ID NO: 26 and VL: SEQ ID NO: 21.
  • the isolated antigen binding protein comprises HCDR3 comprising the amino acid sequence shown in SEQ ID NO:45.
  • the isolated antigen binding protein comprises HCDR2 comprising the amino acid sequence shown in SEQ ID NO:43.
  • the isolated antigen binding protein comprises HCDR1 comprising the amino acid sequence shown in SEQ ID NO:41.
  • the isolated antigen binding protein comprises HCDR1, HCDR2 and HCDR3 of the heavy chain variable region VH shown in SEQ ID NO:77.
  • the isolated antigen binding protein comprises HCDR1, HCDR2 and HCDR3 of the heavy chain variable region VH shown in any one of SEQ ID NO:39 and SEQ ID NO:52.
  • the antigen binding protein of described isolation comprises heavy chain variable region VH
  • described VH comprises described HCDR1, HCDR2 and HCDR3, and described HCDR3 comprises the aminoacid sequence shown in SEQ ID NO:45
  • the HCDR2 comprises the amino acid sequence shown in SEQ ID NO: 43
  • the HCDR1 comprises the amino acid sequence shown in SEQ ID NO: 41.
  • the isolated antigen binding protein comprises H-FR1, the C-terminus of the H-FR1 is directly or indirectly linked to the N-terminus of the HCDR1, and the H-FR1 comprises SEQ ID NO : The amino acid sequence shown in 72.
  • the H-FR1 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:40 and SEQ ID NO:53.
  • the isolated antigen binding protein comprises H-FR2, said H-FR2 is located between said HCDR1 and said HCDR2, and said H-FR2 comprises SEQ ID NO: 73 amino acid sequence.
  • the H-FR2 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:42 and SEQ ID NO:54.
  • said isolated antigen binding protein comprises H-FR3, said H-FR3 is located between said HCDR2 and said HCDR3, and said H-FR3 comprises SEQ ID NO: 74 amino acid sequence.
  • the H-FR3 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:44 and SEQ ID NO:55.
  • the isolated antigen binding protein comprises H-FR4, the N-terminus of the H-FR4 is directly or indirectly linked to the C-terminus of the HCDR3, and the H-FR4 comprises SEQ ID NO : The amino acid sequence shown in 75.
  • the isolated antigen binding protein comprises H-FR4, the N-terminus of the H-FR4 is directly or indirectly linked to the C-terminus of the HCDR3, and the H-FR4 comprises SEQ ID NO :46 and the amino acid sequence shown in any one of SEQ ID NO:30.
  • the isolated antigen-binding protein comprises H-FR1, H-FR2, H-FR3 and H-FR4, and the H-FR1 comprises the amino acid sequence shown in SEQ ID NO: 72; the H-FR2 comprises the amino acid sequence shown in SEQ ID NO: 73; the H-FR3 comprises the amino acid sequence shown in SEQ ID NO: 74; and the H-FR4 comprises the amino acid sequence shown in SEQ ID NO: 75.
  • the H-FR1 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:40 and SEQ ID NO:53;
  • the H-FR2 comprises SEQ ID The amino acid sequence shown in any one of NO:42 and SEQ ID NO:54;
  • the H-FR3 comprises the amino acid sequence shown in any one of SEQ ID NO:44 and SEQ ID NO:55;
  • the H -FR4 comprises the amino acid sequence shown in any one of SEQ ID NO:46 and SEQ ID NO:30.
  • the H-FR1, H-FR2, H-FR3 and H-FR4 in the isolated antigen binding protein comprise an amino acid sequence selected from any of the following groups:
  • H-FR1 SEQ ID NO:40
  • H-FR2 SEQ ID NO:42
  • H-FR3 SEQ ID NO:44
  • H-FR4 SEQ ID NO:46;
  • H-FR1 SEQ ID NO:53
  • H-FR2 SEQ ID NO:54
  • H-FR3 SEQ ID NO:55
  • H-FR4 SEQ ID NO:30.
  • the isolated antigen binding protein comprises a heavy chain variable region VH comprising the amino acid sequence shown in SEQ ID NO:77.
  • the VH in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:39 and SEQ ID NO:52.
  • the isolated antigen binding protein comprises LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 37.
  • the isolated antigen binding protein comprises LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 35.
  • the isolated antigen binding protein comprises LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 33.
  • the isolated antigen binding protein comprises LCDR1, LCDR2 and LCDR3 of the light chain variable region VL shown in SEQ ID NO:76.
  • the isolated antigen binding protein comprises LCDR1, LCDR2 and LCDR3 of the light chain variable region VL shown in any one of SEQ ID NO:31 and SEQ ID NO:47.
  • the antigen binding protein of described separation comprises light chain variable region VL, and described VL comprises described LCDR1, LCDR2 and LCDR3, and described LCDR3 comprises the aminoacid sequence shown in SEQ ID NO:37;
  • the LCDR2 comprises the amino acid sequence shown in SEQ ID NO: 35; and the LCDR1 comprises the amino acid sequence shown in SEQ ID NO: 33.
  • the isolated antigen binding protein comprises L-FR1, the C-terminus of the L-FR1 is directly or indirectly linked to the N-terminus of the LCDR1, and the L-FR1 comprises SEQ ID NO : Amino acid sequence shown in 68.
  • the L-FR1 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:32 and SEQ ID NO:48.
  • said isolated antigen binding protein comprises L-FR2, said L-FR2 is located between said LCDR1 and said LCDR2, and said L-FR2 comprises SEQ ID NO: 69 amino acid sequence.
  • the L-FR2 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:34 and SEQ ID NO:49.
  • said isolated antigen binding protein comprises L-FR3, said L-FR3 is located between said LCDR2 and said LCDR3, and said L-FR3 comprises SEQ ID NO:70 amino acid sequence.
  • the L-FR3 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:36 and SEQ ID NO:50.
  • the isolated antigen binding protein comprises L-FR4, the N-terminus of the L-FR4 is directly or indirectly linked to the C-terminus of the LCDR3, and the L-FR4 comprises SEQ ID NO : The amino acid sequence shown in 71.
  • the L-FR4 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:38 and SEQ ID NO:51.
  • the antigen binding protein of described separation comprises L-FR1, L-FR2, L-FR3 and L-FR4, and described L-FR1 comprises the aminoacid sequence shown in SEQ ID NO: 68; Said L-FR2 comprises the amino acid sequence shown in SEQ ID NO: 69; said L-FR3 comprises the amino acid sequence shown in SEQ ID NO: 70; and said L-FR4 comprises the amino acid sequence shown in SEQ ID NO: 71.
  • the L-FR1 in the isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO:32 and SEQ ID NO:48; the L-FR2 comprises SEQ ID The amino acid sequence shown in any one of NO:34 and SEQ ID NO:49; Said L-FR3 comprises the amino acid sequence shown in any one of SEQ ID NO:36 and SEQ ID NO:50; And said L -FR4 comprises the amino acid sequence shown in any one of SEQ ID NO:38 and SEQ ID NO:51.
  • said L-FR1, L-FR2, L-FR3 and L-FR4 in said isolated antigen binding protein comprise an amino acid sequence selected from any of the following groups:
  • L-FR1 SEQ ID NO:32
  • L-FR2 SEQ ID NO:34
  • L-FR3 SEQ ID NO:36
  • L-FR4 SEQ ID NO:38;
  • L-FR1 SEQ ID NO:48
  • L-FR2 SEQ ID NO:49
  • L-FR3 SEQ ID NO:50
  • L-FR4 SEQ ID NO:51.
  • the isolated antigen binding protein comprises a VL comprising the amino acid sequence shown in SEQ ID NO:76.
  • said VL in said isolated antigen binding protein comprises the amino acid sequence shown in any one of SEQ ID NO: 31 and 47.
  • the isolated antigen binding protein comprises VH and VL comprising an amino acid sequence selected from any of the following groups:
  • VH SEQ ID NO: 39 and VL: SEQ ID NO: 31;
  • VH SEQ ID NO:52 and VL: SEQ ID NO:47.
  • the isolated antigen binding protein comprises a heavy chain constant region, and the heavy chain constant region comprises an IgG-derived constant region or an IgY-derived constant region.
  • said heavy chain constant region in said isolated antibody binding protein comprises an IgG-derived constant region.
  • said heavy chain constant region in said isolated antigen binding protein comprises a constant region derived from IgGl, IgG2, IgG3 or IgG4.
  • the heavy chain constant region in the isolated antigen binding protein comprises the amino acid sequence shown in SEQ ID NO:56.
  • the isolated antigen binding protein comprises a light chain constant region, and the light chain constant region comprises an Ig ⁇ -derived constant region or an Ig ⁇ -derived constant region.
  • said light chain constant region of said isolated antigen binding protein comprises a constant region derived from human Ig ⁇ .
  • the light chain constant region of the isolated antigen binding protein comprises the amino acid sequence shown in SEQ ID NO:57.
  • the isolated antigen binding protein comprises an antibody or antigen binding fragment thereof.
  • the antigen-binding fragment of the isolated antigen-binding protein is selected from the group consisting of Fab, Fab', F(ab)2, Fv fragment, F(ab')2, scFv, di- scFv, VHH and/or dAb.
  • said antibody in said isolated antigen binding protein is selected from the group consisting of monoclonal antibody, single chain antibody, chimeric antibody, humanized antibody and fully human antibody.
  • the application provides one or more polypeptides comprising said isolated antigen binding protein.
  • the application provides one or more immunoconjugates comprising said isolated antigen binding protein or said polypeptide.
  • the present application provides one or more isolated nucleic acid molecules encoding said isolated antigen binding protein, or said polypeptide.
  • the application provides one or more vectors comprising said isolated nucleic acid molecule.
  • the application provides one or more cells comprising said isolated antigen binding protein, said polypeptide, said immunoconjugate, said isolated nucleic acid molecule and/or said carrier.
  • the present application provides a method for preparing the isolated antigen-binding protein or the polypeptide, the method comprising culturing the cells.
  • the present application provides one or more pharmaceutical compositions comprising said isolated antigen binding protein, said polypeptide, said immunoconjugate, said isolated nucleic acid molecule, said carrier, the cells, and/or pharmaceutically acceptable adjuvants and/or excipients.
  • the present application provides a method for detecting or measuring MASP-2, said method comprising using said isolated antigen-binding protein or said polypeptide.
  • the present application provides a detection kit for MASP-2, which comprises the isolated antigen-binding protein or the polypeptide.
  • the present application provides the use of an isolated antigen-binding protein or said polypeptide in the preparation of a kit for detecting the presence and/or content of MASP-2.
  • the present application provides the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell and/or Use of the pharmaceutical composition in the preparation of medicines for preventing and/or treating diseases or conditions.
  • the present application provides the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell and/or the A pharmaceutical composition for preventing, alleviating and/or treating a disease or condition.
  • the present application provides a method for preventing and/or treating a disease or disorder, comprising administering to a subject in need an effective amount of the isolated antigen-binding protein, the polypeptide, the Immunoconjugate, said isolated nucleic acid molecule, said carrier, and/or said cell.
  • Figure 1 shows the binding of an exemplary anti-MASP-2 antibody 50A6 described herein to human MASP-2.
  • Figure 2 shows the binding of an exemplary anti-MASP-2 antibody 47A1 described herein to human MASP-2.
  • Figure 3A shows the binding of an exemplary anti-MASP-2 antibody 50A6 described herein to cynomolgus MASP-2.
  • Figure 3B shows the binding of the exemplary anti-MASP-2 antibody 50A6 described herein to murine MASP-2.
  • Figure 4A shows the binding of an exemplary anti-MASP-2 antibody 47A1 described herein to cynomolgus MASP-2.
  • Figure 4B shows the binding of an exemplary anti-MASP-2 antibody 47A1 described herein to murine MASP-2.
  • Figure 5 shows the results of blocking the lectin pathway of the complement system by exemplary anti-MASP-2 antibodies described herein.
  • Figure 6 shows the results of blocking the lectin pathway of the complement system by an exemplary humanized anti-MASP-2 antibody described in the present application.
  • Figure 7 shows the pharmacokinetic (PK) results of the exemplary anti-MASP-2 antibody described in this application in the humanized FcRn mouse model.
  • isolated generally means obtained from the natural state by artificial means. If an "isolated" substance or component occurs in nature, it may be that its natural environment has been altered, the substance has been isolated from its natural environment, or both. For example, an unisolated polynucleotide or polypeptide naturally exists in a living animal, and the same polynucleotide or polypeptide with high purity isolated from this natural state is called isolation. of.
  • isolated does not exclude the admixture of artificial or synthetic substances, nor the presence of other impure substances which do not affect the activity of the substance.
  • the term “antigen-binding protein” generally refers to a polypeptide molecule capable of specifically recognizing and/or neutralizing a specific antigen.
  • the term “antigen-binding protein” may include “antibody” or "antigen-binding fragment”.
  • the antibody may comprise an immunoglobulin composed of at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, and may include any molecule comprising an antigen-binding portion thereof.
  • antibody may include monoclonal antibodies, antibody fragments or antibody derivatives, including but not limited to murine antibodies, human antibodies (fully human antibodies), humanized antibodies, chimeric antibodies, single chain antibodies (e.g., scFv ), and antibody fragments (eg, Fab, Fab', VHH and (Fab)2 fragments) that bind to the antigen.
  • antibody may also include all recombinant forms of antibodies, such as antibodies expressed in prokaryotic cells, aglycosylated antibodies, and any antigen-binding antibody fragments and derivatives thereof described herein.
  • Each heavy chain can be composed of a heavy chain variable region (VH) and a heavy chain constant region.
  • Each light chain can be composed of a light chain variable region (VL) and a light chain constant region.
  • VH and VL regions can be further distinguished into hypervariable regions called complementarity determining regions (CDRs), which are interspersed in more conserved regions called framework regions (FRs).
  • CDRs complementarity determining regions
  • FRs framework regions
  • Each VH and VL may consist of three CDR and four FR regions, which may be arranged in the following order from amino-terminus to carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.
  • the variable regions of the heavy and light chains contain binding domains that interact with antigen (eg, human MASP-2).
  • the constant regions of the antibodies mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component (Clq) of the classical complement system.
  • the exact boundaries of the CDRs have been defined differently according to different systems.
  • the system described by Kabat Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides a sequence that can be applied to any variable region of an antigen-binding fragment
  • a clear residue numbering system also provides the precise residue boundaries that define the CDRs. These CDRs can be referred to as Kabat CDRs.
  • the term "antigen-binding fragment” generally refers to one or more fragments of an antibody that specifically bind to an antigen.
  • the antigen-binding function of antibodies can be realized by full-length fragments of antibodies.
  • the antigen-binding function of an antibody can also be achieved by comprising a heavy chain of a fragment of Fv, ScFv, dsFv, Fab, Fab' or F(ab'), or by comprising a Fv, scFv, dsFv, Fab, Fab' or The light chain of a fragment of F(ab')2.
  • Fab fragment usually a monovalent fragment consisting of VL, VH, CL and CH domains;
  • F(ab')2 fragment comprising two Fab fragments linked by a disulfide bond at the hinge region (3) Fd fragment composed of VH and CH domains; (4) Fv fragment composed of VL and VH domains of antibody single arm; (5) dAb fragment composed of VH domains (Ward et al., (1989) Nature 341:544-546); (6) an isolated complementarity determining region (CDR) and (7) a combination of two or more isolated CDRs optionally linked by a linker.
  • CDR complementarity determining region
  • the monovalent single-chain molecule Fv formed by the pairing of VL and VH (see Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc.Natl.Acad.Sci.85 :5879-5883).
  • a type of antibody VHH that lacks the light chain of the antibody and only has the variable region of the heavy chain can also be included (for example, see Kang Xiaozhen et al., Acta Biological Engineering, 2018, 34(12): 1974-1984).
  • the "antigen binding portion” may also include an immunoglobulin fusion protein comprising a binding domain selected from: (1) a binding domain polypeptide fused to an immunoglobulin hinge region polypeptide; (2) a binding domain polypeptide fused to an immunoglobulin hinge region polypeptide; an immunoglobulin heavy chain CH2 constant region fused to the hinge region; and (3) an immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region.
  • an immunoglobulin fusion protein comprising a binding domain selected from: (1) a binding domain polypeptide fused to an immunoglobulin hinge region polypeptide; (2) a binding domain polypeptide fused to an immunoglobulin hinge region polypeptide; an immunoglobulin heavy chain CH2 constant region fused to the hinge region; and (3) an immunoglobulin heavy chain CH3 constant region fused to the CH2 constant region.
  • the term "monoclonal antibody” generally refers to a population of substantially homogeneous antibodies, ie, the individual antibodies comprising the population are identical except for possible naturally occurring mutations that are present in minor amounts.
  • Monoclonal antibodies are highly specific, directed against a single antigenic site.
  • the monoclonal antibodies can be produced by hybridoma technology or produced in bacterial, eukaryotic or plant cells by using recombinant DNA methods.
  • Monoclonal antibodies can also be obtained from phage antibody libraries using techniques such as those described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., Mol. Biol., 222:581-597 (1991) conduct.
  • chimeric antibody generally refers to an antibody in which a part of each heavy or light chain amino acid sequence is homologous to the corresponding amino acid sequence in an antibody from a specific species, or belongs to a specific class, and The remainder of the chain is then homologous to the corresponding sequence in another species.
  • the variable regions of both the light and heavy chains are derived from the variable regions of antibodies from one animal species (e.g., mouse, rat, etc.), while the constant portions are homologous to antibody sequences from another species (e.g., human) .
  • B cells or hybridoma cells of non-human origin can be used to produce variable regions combined with constant regions of human origin.
  • variable region has the advantage of being easy to prepare and its specificity is not affected by the source of the constant region it is combined with.
  • the constant region of the chimeric antibody can be derived from humans, the possibility of the chimeric antibody triggering an immune response when injected is lower than that of an antibody whose constant region is of non-human origin.
  • humanized antibody generally refers to a chimeric antibody that contains less sequence from a non-human immunoglobulin, thereby reducing the immunogenicity of a heterologous antibody when introduced into a human, while simultaneously Preserves the full antigen-binding affinity and specificity of the antibody.
  • CDR grafting (Jones et al., Nature 321:522 (1986)) and variants thereof; including “reshaping", (Verhoeyen, et al., 1988 Science 239:1534-1536; Riechmann , et al., 1988 Nature 332:323-337; Tempest, et al., Bio/Technol 1991 9:266-271), "high addition” (hyperchimerization), (Queen, et al., 1989 Proc Natl Acad Sci USA 86:10029-10033; Co, et al., 1991 Proc Natl Acad Sci USA 88:2869-2873; Co, et al., 1992 J Immunol 148:1149-1154) and "veneering", (Mark, et al., "Derivation of therapeutically active humanized and veneered anti-CD18 antibodies.” In: Metcalf B W, Dalton B J, eds.
  • murine antibody generally refers to an antibody whose variable region framework and CDR regions are derived from mouse germline immunoglobulin sequences. In addition, if the antibody comprises a constant region, these also are derived from mouse germline immunoglobulin sequences.
  • the murine antibody of the present application may contain amino acid residues not encoded by mouse germline immunoglobulin sequences, for example, may include mutations introduced by random or point mutations in vitro or by somatic mutations in vivo.
  • germline sequence generally refers to the sequence of unrearranged immunoglobulin DNA sequences.
  • the term "between” usually means that the C-terminal of a certain amino acid fragment is directly or indirectly connected to the N-terminal of the first amino acid fragment, and its N-terminal is directly or indirectly connected to the C-terminal of the second amino acid fragment.
  • indirect connection In the light chain, for example, the N-terminal of the L-FR2 is directly or indirectly connected to the C-terminal of the LCDR1, and the C-terminal of the L-FR2 is directly or indirectly connected to the N-terminal of the LCDR2.
  • the N-terminal of the L-FR3 is directly or indirectly connected to the C-terminal of the LCDR2, and the C-terminal of the L-FR3 is directly or indirectly connected to the N-terminal of the LCDR3.
  • the N-terminal of the H-FR2 is directly or indirectly connected to the C-terminal of the HCDR1
  • the C-terminal of the H-FR2 is directly or indirectly connected to the N-terminal of the HCDR2.
  • the N-terminal of the H-FR3 is directly or indirectly connected to the C-terminal of the HCDR2
  • the C-terminal of the H-FR3 is directly or indirectly connected to the N-terminal of the HCDR3.
  • MASP-2 protein MASP-2
  • MASP-2 antigen any functionally active fragments, variants and homologues of MASP-2, which are derived from Cells express either naturally or on cells transfected with the MASP-2 gene.
  • MASP-2 may be human MASP-2, whose accession number in UniProt/Swiss-Prot is O00187.
  • MASP-2 can be a functionally active fragment of human MASP-2.
  • MASP-2 may be monkey MASP-2 or a functionally active fragment thereof.
  • such “functionally active fragments” may include fragments that retain at least one endogenous function of a naturally occurring protein (eg, binding to an antigen binding protein described herein).
  • the "functionally active fragment” may include a domain that binds to the antigen-binding protein of the present application.
  • MASP-2 is a key regulator of the clusterin pathway of the complement system.
  • Mannan-binding lectin (MBL) or ficolin (FCN) in plasma directly recognizes mannose, N-acetylmannose, N-acetylglucosamine, and rock on the surface of various pathogenic microorganisms. Cocose and the like are sugar structures with terminal sugar groups.
  • the MBL-MASP complex binds to the sugar structure on the surface of the pathogen to activate MASP-1 and MASP-2 independently.
  • Activated MASP-2 exerts its SP activity, cleaves C4, and the resulting C4b fragment is covalently bound to the surface of the pathogen.
  • C2 By interacting with C2, the latter is also cleaved by MASP-2, forming C3 convertase C4b2a, and then activated Complement classical pathway; activated MASP1 can directly cleave C3 to generate C3b, under the action of protein factor D and protein factor P, form C3 convertase C3bBb or C3bBbP, and generate C5 convertase C3bBb3b, activate the alternative pathway of complement.
  • the term "IgA nephropathy” generally refers to primary glomerulopathy of the mesangial region.
  • the "IgA nephropathy” may include primary glomerulopathy mainly characterized by IgA or IgA deposition, with or without deposition of other immunoglobulins in the glomerular mesangium.
  • the lesion types of "IgA nephropathy” may include focal segmental lesions, intracapillary proliferative lesions, mesangial proliferative lesions, crescentic lesions, and sclerosing lesions. Its clinical manifestations are recurrent gross hematuria or microscopic hematuria, which may be accompanied by varying degrees of proteinuria, and some patients may develop severe hypertension or renal insufficiency.
  • the present application may also include functionally active fragments, derivatives, analogs, homologues and fragments thereof.
  • a functionally active fragment refers to a polypeptide having substantially the same amino acid sequence or encoded by a substantially identical nucleotide sequence as the naturally occurring sequence and capable of possessing one or more activities of the naturally occurring sequence.
  • a functionally active fragment of any given sequence is one in which a specific sequence of residues (whether amino acid or nucleotide residues) has been modified such that the polypeptide or polynucleotide substantially retains at least A sequence of endogenous function.
  • Sequences encoding functionally active fragments can be obtained by addition, deletion, substitution, modification, substitution and/or variation of at least one amino acid residue and/or nucleotide residue present in naturally occurring proteins and/or polynucleotides , as long as the original functional activity is maintained.
  • derivative generally refers to the polypeptide or polynucleotide of the present application including any substitution, variation, modification, substitution, deletion and and/or added, so long as the resulting polypeptide or polynucleotide substantially retains at least one of its endogenous functions.
  • analogue generally refers to polypeptides or polynucleotides, including any mimetic of polypeptides or polynucleotides, that is, having at least one endogenous function of the polypeptide or polynucleotide simulated by the mimetic of chemical compounds.
  • amino acid substitutions e.g., at least 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 20) amino acid substitutions can be made so long as the modified sequence remains substantially as desired. activity or ability. Amino acid substitutions may involve the use of non-naturally occurring analogs.
  • homologue generally refers to an amino acid sequence or nucleotide sequence having a certain homology to a naturally occurring sequence.
  • the term “homology” may be equated with sequence "identity”.
  • homologous sequences may include amino acid sequences that may be at least 80%, 85%, 90%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% identical to the subject sequence .
  • a homologue will comprise the same active site, etc., as the subject amino acid sequence.
  • Homology can be considered in terms of similarity (ie, amino acid residues having similar chemical properties/functions), or can be expressed in terms of sequence identity.
  • sequence having a percentage identity of any one of the SEQ ID NOs of the mentioned amino acid sequence or nucleotide sequence means having said percentage identity over the entire length of the mentioned SEQ ID NO the sequence of.
  • sequence alignment can be performed by various means known to those skilled in the art, for example, using BLAST, BLAST-2, ALIGN, NEEDLE or Megalign (DNASTAR) software and the like. Those skilled in the art can determine appropriate parameters for alignment, including any algorithms needed to achieve optimal alignment across the full-length sequences being compared.
  • proteins or polypeptides used in this application may also have deletions, insertions, or substitutions of amino acid residues that produce silent changes and result in functionally equivalent proteins.
  • Deliberate amino acid substitutions can be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or amphipathic nature of the residues, so long as endogenous function is preserved.
  • negatively charged amino acids include aspartic acid and glutamic acid
  • positively charged amino acids include lysine and arginine
  • amino acids with uncharged polar headgroups with similar hydrophilicity values include aspartic acid.
  • Paragine, Glutamine, Serine, Threonine and Tyrosine are examples of amino acid residues that produce silent changes and result in functionally equivalent proteins.
  • the term “immunoconjugate” generally refers to a conjugate formed by conjugating (for example, covalently linking through a linker molecule) the other therapeutic agent to the isolated antigen-binding protein, the conjugate
  • the other therapeutic agent can be delivered to the target cell by specific binding of the isolated antigen binding protein to an antigen on the target cell.
  • the antigen may also be secreted by the target cell and located in the space outside the target cell.
  • subject generally refers to human or non-human animals, including but not limited to cats, dogs, horses, pigs, cows, sheep, rabbits, mice, rats or monkeys.
  • nucleic acid molecule generally refers to an isolated form of nucleotides, deoxyribonucleotides or ribonucleotides or analogs thereof of any length isolated from their natural environment or artificially synthesized.
  • the term "vector” generally refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked.
  • the vectors can transfer inserted nucleic acid molecules into and/or between cells.
  • the vectors may include vectors mainly used for inserting DNA or RNA into cells, vectors mainly used for replicating DNA or RNA, and vectors mainly used for expression of transcription and/or translation of DNA or RNA.
  • the vector may be a polynucleotide capable of being transcribed and translated into a polypeptide when introduced into a suitable cell.
  • the vector produces the desired expression product by culturing appropriate cells containing the vector.
  • the vector may comprise a lentiviral vector.
  • the term "cell” generally refers to an individual cell that can or has contained a plasmid or vector comprising a nucleic acid molecule described herein, or is capable of expressing a polypeptide described herein or an antigen binding protein described herein , cell line or cell culture.
  • the cells may include progeny of a single cell. Due to natural, accidental or deliberate mutations, the progeny cells may not necessarily be completely identical in morphology or genome to the original parent cells, but it is sufficient to be able to express the polypeptide or antigen-binding protein described in this application.
  • the cells can be obtained by transfecting cells in vitro with the vectors described in this application.
  • the cells can be prokaryotic cells (such as Escherichia coli) or eukaryotic cells (such as yeast cells, such as COS cells, Chinese hamster ovary (CHO) cells, HeLa cells, HEK293 cells, COS-1 cells, NSO cells or myeloma cells).
  • the cells can be immune cells.
  • the immune cells may be selected from the group consisting of T cells, B cells, natural killer cells (NK cells), macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, leukocytes and / or peripheral blood mononuclear cells.
  • treating generally refers to: (i) preventing a disease, disorder, and/or condition from occurring in a patient who may be predisposed to it but has not been diagnosed with it; (ii) inhibiting the disease , disorder or condition, i.e. arresting its development; and (iii) relieving the disease, disorder or condition, i.e. making the disease, disorder and/or condition and/or symptoms associated with the disease, disorder and/or condition subside.
  • polypeptide polypeptide
  • peptide protein
  • protein protein
  • proteins are used interchangeably and generally refer to a polymer of amino acids of any length.
  • the polymer can be linear or branched, it can contain modified amino acids, and it can be interrupted by non-amino acids. These terms also encompass amino acid polymers that have been modified. These modifications may include: disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation (such as binding to a labeling component).
  • amino acid includes natural and/or unnatural or synthetic amino acids, including glycine and D and L optical isomers, as well as amino acid analogs and peptidomimetics.
  • polynucleotide used interchangeably and generally refer to nucleosides of any length
  • a polymeric form of an acid such as deoxyribonucleotides or ribonucleotides, or analogs thereof.
  • a polynucleotide can have any three-dimensional structure and can perform any function, known or unknown.
  • polynucleotides coding or non-coding regions of a gene or gene fragment, multiple loci (one locus) defined by junctional analysis, exons, introns, messenger RNA (mRNA), Transfer RNA, ribosomal RNA, short interfering RNA (siRNA), short hairpin RNA (shRNA), micro-RNA (miRNA), ribozyme, cDNA, recombinant polynucleotide, branched polynucleotide, plasmid, vector, any sequence Isolated DNA of any sequence, isolated RNA, nucleic acid probes, and primers.
  • mRNA messenger RNA
  • Transfer RNA Transfer RNA
  • ribosomal RNA short interfering RNA
  • shRNA short hairpin RNA
  • miRNA micro-RNA
  • ribozyme ribozyme
  • cDNA recombinant polynucleotide
  • branched polynucleotide plasmid
  • vector any
  • a polynucleotide may comprise one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs. If present, modification of the nucleotide structure can be performed before or after polymer assembly. The sequence of nucleotides may be interrupted by non-nucleotide components. Polynucleotides can be further modified after polymerization, such as by conjugation with labeled components.
  • K D (likewise, “K D " or “K D ”) generally refers to "affinity constant” or "equilibrium dissociation constant” and refers to a titration measurement at equilibrium, or Value obtained by dividing the dissociation rate constant (kd) by the association rate constant (ka).
  • the association rate constant (ka), dissociation rate constant (kd), and equilibrium dissociation constant ( KD ) are used to express the binding protein (such as the isolated antigen binding protein described herein) to the antigen (such as MASP-2 protein) binding affinity.
  • Methods for determining association and dissociation rate constants are well known in the art. The use of fluorescence-based techniques provides high sensitivity and the ability to examine samples at equilibrium in physiological buffers.
  • the KD value can be determined by Biacore (Biomolecular Interaction Analysis) (for example, an instrument available from BIAcore International AB, a GE Healthcare company, Uppsala, Sweden), and other experimental approaches and instruments such as Octet can also be used.
  • the KD value can also be determined using KinExA (Kinetic Exclusion Assay) available from Sapidyne Instruments (Boise, Idaho), or using a surface plasmon resonance (SPR) instrument.
  • the KD value can also be determined by an amine coupling kit.
  • the term "about” generally refers to a range of 0.5%-10% above or below the specified value, such as 0.5%, 1%, 1.5%, 2%, 2.5%, above or below the specified value. 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10%.
  • the application provides an isolated antigen binding protein, which can have a KD value of about 2E- 09M or less in an Octet assay (for example, the KD is not higher than about 2E- 09M , not higher than About 1.5E-09M, not higher than about 1E-09M, not higher than about 9E-10M, not higher than about 8E-10M, not higher than about 7E-10M, not higher than about 6E-10M, not higher than about 5E-10M, not more than about 2E-10M, not more than about 1E-10M, not more than about 5E-11M, not more than about 1E-11M, or not more than 5E-12M or less) and human MASP-2 Protein-specific binding.
  • the KD is not higher than about 2E- 09M , not higher than About 1.5E-09M, not higher than about 1E-09M, not higher than about 9E-10M, not higher than about 8E-10M, not higher than about 7E-10M, not higher than about 6E-10M, not higher than about 5E
  • the application provides an isolated antigen binding protein, which can have a KD value of about 2E- 09M or less in an Octet assay (for example, the KD is not higher than about 2E- 09M , not higher than About 1.5E-09M, not higher than about 1E-09M, not higher than about 9E-10M, not higher than about 8E-10M, not higher than about 7E-10M, not higher than about 6E-10M, not higher than about 5E-10M, not more than about 2E-10M, not more than about 1E-10M, not more than about 5E-11M, not more than about 1E-11M, or not more than 5E-12M or less) and monkey MASP-2 Protein-specific binding.
  • the KD is not higher than about 2E- 09M , not higher than About 1.5E-09M, not higher than about 1E-09M, not higher than about 9E-10M, not higher than about 8E-10M, not higher than about 7E-10M, not higher than about 6E-10M, not higher than about 5E
  • the application provides an isolated antigen-binding protein, which may comprise at least one CDR in the variable region VH of an antibody heavy chain, and the VH may comprise amino acids shown in SEQ ID NO: 67 or SEQ ID NO: 77 sequence.
  • the VH can comprise the amino acid sequence shown in any one of SEQ ID NO: 13, 26, 39 and 52.
  • the HCDR of the isolated antigen-binding protein can be divided in any form, as long as the VH is identical to the amino acid sequence shown in any one of SEQ ID NO: 13, 26, 39 and 52, it can be divided in any form to obtain All HCDRs can fall within the protection scope of the present application.
  • the CDR of an antibody also known as the complementarity determining region, is part of the variable region.
  • the amino acid residues in this region may make contacts with the antigen or antigenic epitope.
  • Antibody CDRs can be determined by a variety of coding systems, such as CCG, Kabat, Chothia, IMGT, AbM, comprehensive consideration of Kabat/Chothia, etc. These numbering systems are known in the art, see, for example, http://www.bioinf.org.uk/abs/index.html#kabatnum. Those skilled in the art can use different coding systems to determine the CDR region according to the sequence and structure of the antibody. There may be differences in the CDR regions using different coding systems.
  • the CDR covers the CDR sequence divided according to any CDR division method; also covers its variants, the variants include the amino acid sequence of the CDR through substitution, deletion and/or addition of one or more amino acids .
  • the variants include the amino acid sequence of the CDR through substitution, deletion and/or addition of one or more amino acids .
  • amino acids For example 1-30, 1-20 or 1-10, and for example 1, 2, 3, 4, 5, 6, 7, 8 or 9 amino acid substitutions, deletions and/or or insertions; homologues thereof, which may be at least about 85% (e.g., at least about 85%, about 90%, about 91%, about 92%, Amino acid sequences having about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence homology.
  • the isolated antigen binding proteins described herein are defined by the Chothia coding system.
  • the antigen binding protein may comprise a heavy chain variable region VH, and the VH may comprise at least one, two or three of HCDR1, HCDR2 and HCDR3.
  • the HCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 19.
  • the HCDR3 sequence of the antigen binding protein can be defined according to the Chothia coding system.
  • the HCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 17.
  • the HCDR2 sequence of the antigen binding protein can be defined according to the Chothia coding system.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 15.
  • the HCDR1 sequence of the antigen binding protein can be defined according to the Chothia coding system.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 15; the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 17; and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 19 The amino acid sequence shown.
  • the antigen-binding protein may comprise antibody 50A6, JYB1931A63, or an antigen-binding fragment having the same HCDR3 (eg, having the same HCDR1-3) therewith.
  • the VH of the antigen binding protein may comprise framework regions H-FR1, H-FR2, H-FR3 and H-FR4.
  • the H-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:62.
  • H-FR1 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 16 and X19 .
  • EVQLVESGGGLVQPG X 16 SL X 19 LSCAAS (SEQ ID NO: 62), wherein X 16 can be G or R, and X 19 can be R or S.
  • the H-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:14 and SEQ ID NO:27.
  • the H-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:63.
  • H-FR2 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 8 and x10 .
  • NMAWVRQ X 8 P X 10 KGLEWVATI SEQ ID NO: 63
  • X 8 can be A or T
  • X 10 can be G or K.
  • the H-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:16 and SEQ ID NO:28.
  • the H-FR3 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:64.
  • H-FR3 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 16 , X 20 , X 21 , X 27 , X 31 and X 36 .
  • TYYRDSVKGRFTISR X 16 NAK X 20 X 21 LYLQM X 27 SLR X 31 EDTA X 36 YYCST SEQ ID NO: 64, wherein, X 16 can be D or E, X 20 can be N or S, X 21 can be S or T, X 27 can be D or N, X 31 can be A or S, X 36 can be T or V.
  • the H-FR3 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:18 and SEQ ID NO:29.
  • the H-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:65.
  • H-FR4 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 5 and x6 .
  • WGQG X 5 X 6 VTVSS (SEQ ID NO: 65), wherein X 5 can be T or V, and X 6 can be L or M.
  • the H-FR4 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:20 and SEQ ID NO:30.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 62; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 63; the H-FR3 It may comprise the amino acid sequence shown in SEQ ID NO: 64; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 65.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO: 14 and SEQ ID NO: 27; the H-FR2 may comprise SEQ ID NO: 16 and The amino acid sequence shown in any one of SEQ ID NO:28;
  • the H-FR3 can comprise the amino acid sequence shown in any one of SEQ ID NO:18 and SEQ ID NO:29;
  • the H-FR4 can be Comprising the amino acid sequence shown in any one of SEQ ID NO:20 and SEQ ID NO:30.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 14; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 16; the H-FR3 The amino acid sequence shown in SEQ ID NO: 18 may be included; and the H-FR4 may include the amino acid sequence shown in SEQ ID NO: 20.
  • the antigen binding protein may comprise antibody 50A6 or an antigen binding fragment thereof having the same H-FR1-4.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 27; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 28; the H-FR3 It may comprise the amino acid sequence shown in SEQ ID NO: 29; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 30.
  • the antigen-binding protein may include antibody JYB1931A63 or an antigen-binding fragment thereof having the same H-FR1-4.
  • the antigen-binding protein may comprise a heavy chain variable region, and the heavy chain variable region may comprise the amino acid sequence shown in SEQ ID NO:67.
  • the antigen-binding protein comprises a VH having amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group consisting of the sequence shown in SEQ ID NO: 67 compared to the VH: X16 , X19 , X40 , X42 , X73 , X77 , X78 , X84 , X88 , X93 , X119 , X120 .
  • X 40 can be A or T
  • X 42 can be G or K
  • X 73 can be D or E
  • X 77 can be N or S
  • X 78 can be S or T
  • X 84 can be D or N
  • X 88 can be A or S
  • X 93 can be T or V
  • X 119 can be T or V
  • X 120 can be L or M.
  • the heavy chain variable region of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:13 and SEQ ID NO:26.
  • the antigen-binding protein may comprise a heavy chain constant region, which may include an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:56.
  • the antigen-binding protein may comprise at least one CDR in the variable region VL of the antibody light chain, and the VL may comprise the amino acid sequence shown in SEQ ID NO: 66 or SEQ ID NO: 76.
  • the VL may comprise the amino acid sequence shown in any one of SEQ ID NO:66 or SEQ ID NO:76.
  • the LCDR of the isolated antigen-binding protein can be divided in any form, as long as the VL is identical to the amino acid sequence shown in any one of SEQ ID NO: 66 or SEQ ID NO: 76, it can be divided in any form All LCDRs can fall within the protection scope of the present application.
  • the antigen binding protein may comprise a light chain variable region VL, and the VL may comprise at least one, at least two or at least three of LCDR1, LCDR2 and LCDR3.
  • the LCDR3 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO: 11.
  • the LCDR3 of the antigen binding protein can be defined according to the Chothia numbering system.
  • the LCDR2 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO: 9.
  • the LCDR2 of the antigen binding protein can be defined according to the Chothia numbering system.
  • the LCDR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:7.
  • the LCDR1 of the antigen binding protein can be defined according to the Chothia numbering system.
  • the LCDR1 of the antigen binding protein described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 7; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 9; and the LCDR3 may comprise SEQ ID NO: The amino acid sequence shown in 11.
  • the antigen binding protein may comprise antibody 50A6, JYB1931A63, or an antigen binding fragment having the same LCDR3 (eg, having the same LCDR1-3) therewith.
  • the VL of the antigen binding protein may comprise the framework regions L-FR1, L-FR2, L-FR3 and L-FR4.
  • the L-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:58.
  • L-FR1 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the following group: X 1 , X 13 , X 14 , X 18 , X 19 and X 20 .
  • X 1 IVLTQSPATLS X 13 X 14 PGE X 18 X 19 X 20 LSC (SEQ ID NO: 58), wherein X 1 can be E or N, X 13 can be L or V, X 14 can be S or T, X 18 can be R or S, X 19 can be A or V, X 20 can be S or T.
  • the L-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:6 and SEQ ID NO:22.
  • the L-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:59.
  • L-FR2 of the antigen-binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 5 , X 6 , X 7 , X 8 and X 9 .
  • WYQQ X 5 X 6 X 7 X 8 X 9 PRLLIK (SEQ ID NO: 59), wherein X 5 can be K or R, X 6 can be P or S, X 7 can be G or N, and X 8 can be E or Q, X9 can be A or S.
  • the L-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:8 and SEQ ID NO:23.
  • the L-FR3 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:60.
  • L-FR3 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 4 , X 18 , X 20 , X 21 , X 22 , X 24 , X 28 and X 29 .
  • GIPX 4 RFSGSGSGTDFTLX 18 I X 20 X 21 X 22 EX 24 EDFX 28 X 29 YYC (SEQ ID NO: 60), wherein X 4 can be A or S, X 18 can be S or T, and X 20 can be N or S , X 21 can be R or S, X 22 can be L or V, X 24 can be P or S, X 28 can be A or S, and X 29 can be I or V.
  • the L-FR3 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:10 and SEQ ID NO:24.
  • the L-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:61.
  • L-FR4 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 3 , X 9 and X 10 .
  • FG X 3 GTKLE X 9 X 10 (SEQ ID NO: 61), wherein X 3 can be A or Q, X 9 can be I or L, and X 10 can be K or R.
  • the L-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:12 and SEQ ID NO:25.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 58; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 59; the L-FR3 It may comprise the amino acid sequence shown in SEQ ID NO: 60; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 61.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:6 and SEQ ID NO:22; the L-FR2 may comprise SEQ ID NO:8 and The amino acid sequence shown in any one of SEQ ID NO:23;
  • the L-FR3 can comprise the amino acid sequence shown in any one of SEQ ID NO:10 and SEQ ID NO:24;
  • the L-FR4 can be Comprising the amino acid sequence shown in any one of SEQ ID NO:12 and SEQ ID NO:25.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 6; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 8; the L-FR3 The amino acid sequence shown in SEQ ID NO: 10 may be included; and the L-FR4 may include the amino acid sequence shown in SEQ ID NO: 12.
  • the antigen binding protein may comprise antibody 50A6 or an antibody having the same L-FR1-4 as it.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 22; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 23; the L-FR3 It may comprise the amino acid sequence shown in SEQ ID NO: 24; and the L-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 25.
  • the antigen-binding protein may include antibody JYB1931A63 or an antibody having the same H-FR1-4 as it.
  • the antigen-binding protein may comprise a light chain variable region VL, and the VL may comprise the amino acid sequence shown in SEQ ID NO:66.
  • the VL of the antigen-binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 1 , X 13 , X 14 , X 18 , X 19 , X 20 , X 39 , X 40 , X 41 , X 42 , X 43 , X 60 , X 74 , X 76 , X 77 , X 78 , X 80 , X 84 , X 85 , X 100 , X 106 , X 107 .
  • the light chain variable region of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:5 and SEQ ID NO:21.
  • the antigen-binding protein may comprise a light chain constant region, which may include a constant region derived from Ig ⁇ or a constant region derived from Ig ⁇ .
  • the light chain constant region may comprise a constant region derived from Ig ⁇ .
  • the light chain constant region of the antigen binding protein comprises the amino acid sequence shown in SEQ ID NO:57.
  • the antigen binding protein may comprise HCDR1-3 and LCDR1-3.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 15; the HCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 17; the HCDR3 of the antigen binding protein may comprise Comprising the amino acid sequence shown in SEQ ID NO:19; the LCDR1 of the antigen-binding protein may include the amino acid sequence shown in SEQ ID NO:7; the LCDR2 of the antigen-binding protein may include the amino acid shown in SEQ ID NO:9 Sequence; the LCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:11.
  • the antigen binding protein may comprise antibody 50A6, JYB1931A63, or an antigen binding fragment that has the same HCDR3 (eg, has the same HCDR1-3) and LCDR3 (eg, has the same LCDR1-3) thereof.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the antigen binding protein may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region of the antigen binding protein may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO:15;
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:17;
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:19;
  • the LCDR1 can include the amino acid sequence shown in SEQ ID NO:7;
  • the LCDR2 can include the amino acid sequence shown in SEQ ID NO:9;
  • the LCDR3 can include the amino acid sequence shown in SEQ ID NO:11.
  • the L-FR3 can include the amino acid sequence shown in SEQ ID NO: 10;
  • the L-FR4 can include the amino acid sequence shown in SEQ ID NO: 12.
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 13.
  • the antigen binding protein may comprise antibody 50A6 or an antigen binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:5.
  • the antigen binding protein may comprise antibody 50A6 or an antigen binding protein having the same light chain variable region as it.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the LCDR1 can include the amino acid sequence shown in SEQ ID NO:7; the LCDR2 can include the amino acid sequence shown in SEQ ID NO:9; the LCDR3 can include the amino acid sequence shown in SEQ ID NO:11.
  • the H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:27; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:28; the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:29
  • the amino acid sequence shown; the H-FR4 can comprise the amino acid sequence shown in SEQ ID NO:30; the L-FR1 can comprise the amino acid sequence of SEQ ID NO:22; the L-FR2 can comprise the amino acid sequence of SEQ ID NO The amino acid sequence shown in: 23;
  • the L-FR3 can comprise the amino acid sequence shown in SEQ ID NO: 24;
  • the L-FR4 can comprise the amino acid sequence shown in SEQ ID NO: 25.
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:26.
  • the antigen-binding protein may include antibody JYB1931A63 or an antigen-binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:21.
  • the antigen-binding protein may include antibody JYB1931A63 or an antigen-binding protein having the same light chain variable region as it.
  • the antigen binding protein may comprise a heavy chain variable region VH, and the VH may comprise at least one, two or three of HCDR1, HCDR2 and HCDR3.
  • the HCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 45.
  • the HCDR3 sequence of the antigen binding protein can be defined according to the Chothia coding system.
  • the HCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 43.
  • the HCDR2 sequence of the antigen binding protein can be defined according to the Chothia coding system.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 41.
  • the HCDR1 sequence of the antigen binding protein can be defined according to the Chothia coding system.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 41; the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 43; and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 45.
  • the antigen-binding protein may comprise antibody 47A1, JYB1931A13, or an antigen-binding fragment having the same HCDR3 (eg, having the same HCDR1-3) therewith.
  • the VH of the antigen binding protein may comprise framework regions H-FR1, H-FR2, H-FR3 and H-FR4.
  • the H-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:72.
  • H-FR1 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 1 , X 17 and X 25 .
  • X 1 VQLQESGPGLVKPSQ X 17 LSLTCTV X 25 (SEQ ID NO: 72), wherein X 1 can be D or Q, X 17 can be S or T, and X 25 can be S or T.
  • the H-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:40 and SEQ ID NO:53.
  • the H-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:73.
  • H-FR2 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 8 , X 11 , X 12 and X 16 .
  • AWNWIRQ X 8 PG X 11 X 12 LEW X 16 GYI (SEQ ID NO: 73), wherein X 8 can be F or P, X 11 can be K or N, X 12 can be G or K, and X 16 can be I or M.
  • the H-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:42 and SEQ ID NO:54.
  • the H-FR3 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:74.
  • H-FR3 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the following group: X 11 , X 12 , X 14 , X 16 , X 19 , X 23 , X 25 , X 27 , X 31 , X 32 , X 36 and X 38 .
  • X 11 can be I or V
  • X 12 Can be S or T
  • X 14 can be S or T
  • X 16 can be D or N
  • X 19 can be K or T
  • X 23 can be F or S
  • X 25 can be K or Q
  • X 27 can be N or S
  • X 31 can be A or T
  • X 32 can be A or E
  • X 36 can be T or V
  • X 38 can be F or Y.
  • the H-FR3 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:44 and SEQ ID NO:55.
  • the H-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:75.
  • H-FR4 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 6 and X7 .
  • WGQGT X 6 X 7 TVSS (SEQ ID NO: 75), wherein X 6 can be L or T, and X 7 can be L or V.
  • the H-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:46 and SEQ ID NO:30.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 72; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 73; the H-FR3 It may comprise the amino acid sequence shown in SEQ ID NO: 74; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 75.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:40 and SEQ ID NO:53; the H-FR2 may comprise SEQ ID NO:42 and The amino acid sequence shown in any one of SEQ ID NO:54;
  • the H-FR3 can comprise the amino acid sequence shown in any one of SEQ ID NO:44 and SEQ ID NO:55;
  • the H-FR4 can Comprising the amino acid sequence shown in any one of SEQ ID NO:46 and SEQ ID NO:30.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 40; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 42; the H-FR3 It may comprise the amino acid sequence shown in SEQ ID NO: 44; and the H-FR4 may comprise the amino acid sequence shown in SEQ ID NO: 46.
  • the antigen binding protein may comprise antibody 47A1 or an antigen binding fragment thereof having the same H-FR1-4.
  • the H-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 53; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 54; the H-FR3 The amino acid sequence shown in SEQ ID NO: 55 may be included; and the H-FR4 may include the amino acid sequence shown in SEQ ID NO: 30.
  • the antigen-binding protein may include antibody JYB1931A13 or an antigen-binding fragment thereof having the same H-FR1-4.
  • the antigen-binding protein may comprise a heavy chain variable region, and the heavy chain variable region may comprise the amino acid sequence shown in SEQ ID NO:77.
  • the antigen-binding protein comprises a VH having amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group consisting of the sequence shown in SEQ ID NO: 77: X1, X17 , X25 , X41 , X44 , X45 , X49, X68, X69 , X71 , X73 , X76 , X80 , X82 , X84 , X88 , X89 , X 93 , X 95 , X 109 and X 110 .
  • the heavy chain variable region of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:39 and SEQ ID NO:52.
  • the antigen-binding protein may comprise a heavy chain constant region, which may include an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:56.
  • the antigen binding protein may comprise a light chain variable region VL, and the VL may comprise at least one, at least two or at least three of LCDR1, LCDR2 and LCDR3.
  • the LCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 37.
  • the LCDR3 of the antigen binding protein can be defined according to the Chothia numbering system.
  • the LCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 35.
  • the LCDR2 of the antigen binding protein can be defined according to the Chothia numbering system.
  • the LCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 33.
  • the LCDR1 of the antigen binding protein can be defined according to the Chothia numbering system.
  • the LCDR1 of the antigen binding protein described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 33; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 35; and the LCDR3 may comprise SEQ ID NO: The amino acid sequence shown in 37.
  • the antigen binding protein may comprise antibody 47A1, JYB1931A13, or an antigen binding fragment having the same LCDR3 (eg, having the same LCDR1-3) therewith.
  • the VL of the antigen binding protein may comprise the framework regions L-FR1, L-FR2, L-FR3 and L-FR4.
  • the L-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:68.
  • L-FR1 of the antigen-binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 3 , X 8 , X 9 , X 10 , X 11 , X 13 , X 20 and X 21 .
  • X 3 can be Q or V
  • X 8 can be P or Q
  • X 9 can be K or S
  • X 10 can be F or S
  • X 11 can be L or M
  • X 13 can be A or T
  • X 20 can be S or T
  • X 21 can be I or V.
  • the L-FR1 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:32 and SEQ ID NO:48.
  • the L-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:69.
  • the L-FR2 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 8 and X9 .
  • WFQQKPG X 8 X 9 PKPLIY (SEQ ID NO: 69), wherein X 8 can be K or Q, and X 9 can be A or S.
  • the L-FR2 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:34 and SEQ ID NO:49.
  • the L-FR3 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:70.
  • L-FR3 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 4 , X 7 , X 21 , X 22 , X 24 , X 27 and X 29 .
  • the L-FR3 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:36 and SEQ ID NO:50.
  • the L-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in SEQ ID NO:71.
  • L-FR4 of the antigen binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 3 , X 7 , X 9 , X 10 .
  • X 3 can be A or G
  • X 7 can be L or V
  • X 9 can be I or L
  • X 10 can be K or N.
  • the L-FR4 of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:38 and SEQ ID NO:51.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 68; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 69; the L-FR3 The amino acid sequence shown in SEQ ID NO: 70 may be included; and the L-FR4 may include the amino acid sequence shown in SEQ ID NO: 71.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO:32 and SEQ ID NO:48; the L-FR2 may comprise SEQ ID NO:34 and The amino acid sequence shown in any one of SEQ ID NO:49;
  • the L-FR3 can comprise the amino acid sequence shown in any one of SEQ ID NO:36 and SEQ ID NO:50;
  • the L-FR4 can be Comprising the amino acid sequence shown in any one of SEQ ID NO:38 and SEQ ID NO:51.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 32; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 34; the L-FR3 The amino acid sequence shown in SEQ ID NO: 36 can be included; and the L-FR4 can include the amino acid sequence shown in SEQ ID NO: 38.
  • the antigen binding protein may include antibody 47A1 or an antibody having the same L-FR1-4 as it.
  • the L-FR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 48; the L-FR2 may comprise the amino acid sequence shown in SEQ ID NO: 49; the L-FR3 The amino acid sequence shown in SEQ ID NO: 50 may be included; and the L-FR4 may include the amino acid sequence shown in SEQ ID NO: 51.
  • the antigen binding protein may include antibody JYB1931A13 or an antibody having the same H-FR1-4 as it.
  • the antigen-binding protein may comprise a light chain variable region VL, and the VL may comprise the amino acid sequence shown in SEQ ID NO:76.
  • the VL of the antigen-binding protein has amino acid substitutions (eg, conservative amino acid substitutions, etc.) at one or more amino acids selected from the group: X 3 , X 8 , X 9 , X 10 , X 11 , X 13 , X 20 , X 21 , X 42 , X 43 , X 60 , X 63 , X 77 , X 78 , X 80 , X 83 , X 85 , X 100 , X 104 , X 106 , X 107 .
  • X 3 can be Q or V
  • X 8 can be P or Q
  • X 9 can be K or S
  • X 10 can be F or S
  • X 11 can be L or M
  • X 13 can be A or T
  • X 20 can be S or T
  • X 21 can be I or V
  • X 42 can be K or Q
  • X 43 can be A or S
  • X 60 can be D or S
  • X 63 can be S or T
  • X 77 can be N or S
  • X 78 can be L
  • the light chain variable region of the antigen-binding protein may comprise the amino acid sequence shown in any one of SEQ ID NO: 31 and 47.
  • the antigen-binding protein may comprise a light chain constant region, which may include a constant region derived from Ig ⁇ or a constant region derived from Ig ⁇ .
  • the light chain constant region may comprise a constant region derived from Ig ⁇ .
  • the light chain constant region of the antigen binding protein comprises the amino acid sequence shown in SEQ ID NO:57.
  • the antigen binding protein may comprise HCDR1-3 and LCDR1-3.
  • the HCDR1 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:41; the HCDR2 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:43; the HCDR3 of the antigen binding protein may comprise Comprising the amino acid sequence shown in SEQ ID NO:45; the LCDR1 of the antigen-binding protein may include the amino acid sequence shown in SEQ ID NO:33; the LCDR2 of the antigen-binding protein may include the amino acid shown in SEQ ID NO:35 Sequence; the LCDR3 of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:37.
  • the antigen binding protein may comprise antibody 47A1, JYB1931A13 or an antigen binding fragment having the same HCDR3 (eg, having the same HCDR1-3) and LCDR3 (eg, having the same LCDR1-3) thereof.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the antigen binding protein may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region of the antigen binding protein may comprise LCDR1-3 and L-FR1-4.
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO:41;
  • the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:43;
  • the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:45;
  • the LCDR1 can include the amino acid sequence shown in SEQ ID NO:33;
  • the LCDR2 can include the amino acid sequence shown in SEQ ID NO:35;
  • the LCDR3 can include the amino acid sequence shown in SEQ ID NO:37.
  • the L-FR3 can comprise the amino acid sequence shown in SEQ ID NO: 36;
  • the L-FR4 can comprise the amino acid sequence shown in SEQ ID NO: 38.
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:39.
  • the antigen binding protein may comprise antibody 47A1 or an antigen binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:31.
  • the antigen binding protein may comprise antibody 47A1 or an antigen binding protein having the same light chain variable region as it.
  • the antigen binding protein may comprise a heavy chain variable region and a light chain variable region, and the heavy chain variable region may comprise HCDR1-3 and H-FR1-4.
  • the light chain variable region may comprise LCDR1-3 and L-FR1-4.
  • the LCDR1 can include the amino acid sequence shown in SEQ ID NO:33; the LCDR2 can include the amino acid sequence shown in SEQ ID NO:35; the LCDR3 can include the amino acid sequence shown in SEQ ID NO:37.
  • the H-FR1 may comprise the amino acid sequence shown in SEQ ID NO:53; the H-FR2 may comprise the amino acid sequence shown in SEQ ID NO:54; the H-FR3 may comprise the amino acid sequence shown in SEQ ID NO:55
  • the amino acid sequence shown; the H-FR4 can comprise the amino acid sequence shown in SEQ ID NO:30; the L-FR1 can comprise the amino acid sequence of SEQ ID NO:48; the L-FR2 can comprise the amino acid sequence of SEQ ID NO The amino acid sequence shown in: 49;
  • the L-FR3 can comprise the amino acid sequence shown in SEQ ID NO: 50;
  • the L-FR4 can comprise the amino acid sequence shown in SEQ ID NO: 51.
  • the heavy chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO:52.
  • the antigen-binding protein may include antibody JYB1931A13 or an antigen-binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the antigen binding protein may comprise the amino acid sequence shown in SEQ ID NO: 47.
  • the antigen-binding protein may include antibody JYB1931A13 or an antigen-binding protein having the same light chain variable region as it.
  • the isolated antigen-binding protein can also compete with a reference antibody for binding to the human MASP-2 protein, and the reference antibody can comprise a heavy chain variable region VH, and the VH can comprise HCDR1, HCDR2 and at least one, two or three of HCDR3.
  • the HCDR3 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 19.
  • the sequence of HCDR3 of the reference antibody can be defined according to the Chothia coding system.
  • the HCDR2 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 17.
  • the sequence of HCDR2 of the reference antibody can be defined according to the Chothia coding system.
  • the HCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 15.
  • the sequence of HCDR1 of the reference antibody can be defined according to the Chothia coding system.
  • the HCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 15; the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 17; and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 19 The amino acid sequence shown.
  • the reference antibody can include antibody 50A6, JYB1931A63, or an antigen binding protein that has the same HCDR3 (eg, has the same HCDR1-3) as it.
  • the reference antibody may comprise a heavy chain variable region, and the heavy chain variable region may comprise the amino acid sequence shown in SEQ ID NO:67.
  • X 40 can be A or T
  • X 42 can be G or K
  • X 73 can be D or E
  • X 77 can be N or S
  • X 78 can be S or T
  • X 84 can be D or N
  • X 88 can be A or S
  • X 93 can be T or V
  • X 119 can be T or V
  • X 120 can be L or M.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NO:13 and SEQ ID NO:26.
  • the reference antibody may comprise a heavy chain constant region, which may comprise an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:56.
  • the reference antibody may comprise a light chain variable region VL, which may comprise LCDR1, LCDR2 and LCDR3.
  • the LCDR3 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 11.
  • the sequence of the LCDR3 of the reference antibody can be defined according to the Chothia coding system.
  • the LCDR2 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO:9.
  • the sequence of LCDR2 of the reference antibody can be defined according to the Chothia coding system.
  • the LCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO:7.
  • the sequence of LCDR1 of the reference antibody can be defined according to the Chothia coding system.
  • the LCDR1 of the reference antibody described in the present application may comprise the amino acid sequence shown in SEQ ID NO: 7; the LCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 9; and the LCDR3 may comprise SEQ ID NO: The amino acid sequence shown in 11.
  • the reference antibody can include antibody 50A6, JYB1931A63, or an antigen binding protein that has the same LCDR3 (eg, has the same LCDR1-3) therewith.
  • the reference antibody may comprise a light chain variable region, and the light chain variable region may comprise the amino acid sequence shown in SEQ ID NO:66.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NO:5 and SEQ ID NO:21.
  • the reference antibody may comprise HCDR1-3 and LCDR1-3.
  • the reference antibody can include antibody 50A6, JYB1931A63, or an antigen binding protein that has the same HCDR3 (e.g., has the same HCDR1-3) and LCDR3 (e.g., has the same LCDR1-3) as it.
  • the isolated antigen-binding protein can also compete with a reference antibody for binding to the human MASP-2 protein, and the reference antibody can comprise a heavy chain variable region VH, and the VH can comprise HCDR1, HCDR2 and at least one, two or three of HCDR3.
  • the HCDR3 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 45.
  • the sequence of HCDR3 of the reference antibody can be defined according to the Chothia coding system.
  • the HCDR2 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 43.
  • the sequence of HCDR2 of the reference antibody can be defined according to the Chothia coding system.
  • the HCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 41.
  • the sequence of HCDR1 of the reference antibody can be defined according to the Chothia coding system.
  • the HCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 41; the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO: 43; and the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO: 45.
  • the reference antibody may comprise antibody 47A1, JYB1931A13, or an antigen binding protein having the same HCDR3 (eg, having the same HCDR1-3) therewith.
  • the reference antibody may comprise a heavy chain variable region, and the heavy chain variable region may comprise the amino acid sequence shown in SEQ ID NO:77.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NO:39 and SEQ ID NO:52.
  • the reference antibody may comprise a heavy chain constant region, which may comprise an IgG-derived constant region or an IgY-derived constant region.
  • the heavy chain constant region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:56.
  • the reference antibody may comprise a light chain variable region VL, which may comprise LCDR1, LCDR2 and LCDR3.
  • the LCDR3 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 37.
  • the sequence of the LCDR3 of the reference antibody can be defined according to the Chothia coding system.
  • the LCDR2 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 35.
  • the sequence of LCDR2 of the reference antibody can be defined according to the Chothia coding system.
  • the LCDR1 of the reference antibody may comprise the amino acid sequence shown in SEQ ID NO: 33.
  • the sequence of LCDR1 of the reference antibody can be defined according to the Chothia coding system.
  • the reference antibody can include antibody 47A1, JYB1931A13, or an antigen binding protein with the same LCDR3 (eg, with the same LCDR1-3) therewith.
  • the reference antibody may comprise a light chain variable region, and the light chain variable region may comprise the amino acid sequence shown in SEQ ID NO:76.
  • X 3 can be Q or V
  • X 8 can be P or Q
  • X 9 can be K or S
  • X 10 can be F or S
  • X 11 can be L or M
  • X 13 can be A or T
  • X 20 can be S or T
  • X 21 can be I or V
  • X 42 can be K or Q
  • X 43 can be A or S
  • X 60 can be D or S
  • X 63 can be S or T
  • X 77 can be N or S
  • X 78 can be L
  • the light chain variable region of the reference antibody may comprise the amino acid sequence shown in any one of SEQ ID NO: 31 and 47.
  • the reference antibody may comprise HCDR1-3 and LCDR1-3.
  • the HCDR1 may comprise the amino acid sequence shown in SEQ ID NO:41; the HCDR2 may comprise the amino acid sequence shown in SEQ ID NO:43; the HCDR3 may comprise the amino acid sequence shown in SEQ ID NO:45;
  • the LCDR1 can include the amino acid sequence shown in SEQ ID NO:33; the LCDR2 can include the amino acid sequence shown in SEQ ID NO:35; the LCDR3 can include the amino acid sequence shown in SEQ ID NO:37.
  • the reference antibody can include antibody 47A1, JYB1931A13, or an antigen binding protein that has the same HCDR3 (e.g., has the same HCDR1-3) and LCDR3 (e.g., has the same LCDR1-3) as it.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO: 13.
  • the reference antibody can comprise antibody 50A6 or an antigen binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:5.
  • the reference antibody may comprise antibody 50A6 or an antigen binding protein having the same light chain variable region as it.
  • the reference antibody can include antibody 50A6 or an antigen binding protein having the same heavy and light chain variable regions as it.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:26.
  • the reference antibody may include antibody JYB1931A63 or an antigen binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the reference antibody can comprise the amino acid sequence set forth in SEQ ID NO:21.
  • the reference antibody may include antibody JYB1931A63 or an antigen binding protein having the same light chain variable region as it.
  • the reference antibody may include antibody JYB1931A63 or an antigen-binding protein having the same heavy chain variable region and light chain variable region as it.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody can comprise the amino acid sequence set forth in SEQ ID NO:39.
  • the reference antibody can include antibody 47A1 or an antigen binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:31.
  • the reference antibody can comprise antibody 47A1 or an antigen binding protein having the same light chain variable region as it.
  • the reference antibody may comprise antibody 47A1 or an antigen binding protein having the same heavy and light chain variable regions as antibody 47A1.
  • the reference antibody may comprise a heavy chain variable region and a light chain variable region.
  • the heavy chain variable region of the reference antibody may comprise the amino acid sequence set forth in SEQ ID NO:52.
  • the reference antibody may include antibody JYB1931A13 or an antigen binding protein having the same heavy chain variable region as it.
  • the light chain variable region of the reference antibody can comprise the amino acid sequence set forth in SEQ ID NO:47.
  • the reference antibody may include antibody JYB1931A13 or an antigen binding protein having the same light chain variable region as it.
  • the reference antibody may include antibody JYB1931A13 or an antigen-binding protein having the same heavy chain variable region and light chain variable region as it.
  • the application provides one or more polypeptides, which may comprise an isolated antigen binding protein of the application.
  • the polypeptide can include a fusion protein.
  • the polypeptides can include multispecific antibodies (eg, bispecific antibodies).
  • the application provides one or more immunoconjugates, which may comprise an isolated antigen binding protein of the application.
  • the immunoconjugate may further comprise a pharmaceutically acceptable therapeutic agent, label and/or detection agent.
  • the present application also provides one or more isolated nucleic acid molecules that encode the isolated antigen-binding protein described herein.
  • each of the one or more nucleic acid molecules may encode the entirety of the antigen binding protein, or may encode a portion thereof (e.g., HCDR1-3, one of the heavy chain variable regions, or variety).
  • the products encoded by the nucleic acid molecules together can form a functional (eg, bind MASP-2) isolated antigen binding protein of the present application.
  • the nucleic acid molecules described herein can be isolated. For example, it may be produced or synthesized by (i) amplified in vitro, such as by polymerase chain reaction (PCR) amplification, (ii) recombinantly produced by cloning, (iii) purified (iv) synthetic, for example by chemical synthesis.
  • the isolated nucleic acid can be a nucleic acid molecule prepared by recombinant DNA techniques.
  • nucleic acid encoding the isolated antigen-binding protein can be prepared by various methods known in the art, including but not limited to, using reverse transcription PCR and PCR to obtain the isolated antigen-binding protein described in the application. protein nucleic acid molecule.
  • the present application provides one or more vectors comprising one or more nucleic acid molecules described herein.
  • Each vector may contain one or more such nucleic acid molecules.
  • other genes may be included in the vector, such as marker genes that allow selection of the vector in appropriate host cells and under appropriate conditions.
  • the vector may also contain expression control elements that permit proper expression of the coding region in an appropriate host.
  • control elements are well known to those skilled in the art, and may include, for example, promoters, ribosome binding sites, enhancers, and other control elements that regulate gene transcription or mRNA translation, and the like.
  • the expression control sequences are regulatable elements.
  • the specific structure of the expression control sequence may vary depending on the function of the species or cell type, but generally includes 5' non-transcribed sequences and 5' and 3' non-translated sequences involved in the initiation of transcription and translation, respectively, such as TATA box, plus Cap sequence, CAAT sequence, etc.
  • the 5' non-transcribed expression control sequence may comprise a promoter region which may comprise a promoter sequence for transcriptional control of the functionally linked nucleic acid.
  • the expression control sequences may also include enhancer sequences or upstream activator sequences.
  • suitable promoters may include, for example, promoters for SP6, T3, and T7 polymerases, human U6 RNA promoters, CMV promoters, and artificial hybrid promoters thereof (such as CMV), wherein the promoter's Portions may be fused to portions of gene promoters of other cellular proteins (eg, human GAPDH, glyceraldehyde-3-phosphate dehydrogenase), which may or may not contain additional introns.
  • One or more nucleic acid molecules described herein can be operably linked to the expression control element.
  • Such vectors may include, for example, plasmids, cosmids, viruses, phages, or other vectors commonly used in, for example, genetic engineering.
  • the vector may be an expression vector.
  • the vector can be a viral vector.
  • Viral vectors may be administered directly to the patient (in vivo) or may be indirect, for example, in vitro by treating cells with virus and then administering the treated cells to the patient (ex vivo).
  • Viral vector technology is well known in the art and described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York) and other handbooks of virology and molecular biology.
  • Lentiviral vectors are retroviral vectors capable of transducing or infecting non-dividing cells and typically producing higher viral titers.
  • Lentiviral vectors may contain long terminal repeat 5'LTR and truncated 3'LTR, RRE, rev response element (cPPT), central termination sequence (CTS) and/or post-translational regulatory element (WPRE).
  • the vectors described herein can be introduced into cells.
  • the present application provides a cell.
  • the cells may comprise the isolated antigen binding protein described herein, the polypeptide, the immunoconjugate, one or more nucleic acid molecules and/or one or more carriers described herein .
  • each or each cell may comprise one or more of the nucleic acid molecules or vectors described herein.
  • each or each cell may contain multiple (eg, 2 or more) or multiple (eg, 2 or more) nucleic acid molecules or vectors described herein.
  • the vectors described herein can be introduced into said host cells, such as prokaryotic cells (e.g., bacterial cells), CHO cells, NS/0 cells, HEK293T cells, 293F cells or HEK293A cells, or other eukaryotic cells, Such as cells from plants, fungal or yeast cells, etc.
  • the vectors described in this application can be introduced into the host cells by methods known in the art, such as electroporation, lipofectine transfection, lipofectamin transfection and the like.
  • the cells can include yeast cells.
  • the cells may include E. coli cells.
  • the cells can include mammalian cells.
  • the cells can include immune cells.
  • the cells may include immune cells.
  • the cells may include immune cells.
  • the cells may include T cells, B cells, natural killer (NK) cells, macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, leukocytes, and/or peripheral blood mononuclear cells cell.
  • NK natural killer
  • the present application provides a pharmaceutical composition.
  • the pharmaceutical composition may comprise the isolated antigen binding protein described herein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, the cell, and/or Or pharmaceutically acceptable adjuvants and/or excipients.
  • the pharmaceutically acceptable adjuvants may include buffers, antioxidants, preservatives, low molecular weight polypeptides, proteins, hydrophilic polymers, amino acids, sugars, chelating agents, counter ions, metal complexes and /or nonionic surfactants. Unless incompatible with the cells described herein, any conventional media or reagents are contemplated for use in the pharmaceutical compositions of the present application.
  • the pharmaceutically acceptable excipients may include additives other than the main drug in the pharmaceutical preparation, and may also be referred to as auxiliary materials.
  • the excipients may include binders, fillers, disintegrants, lubricants in tablets.
  • the excipients may include wine, vinegar, medicinal juice, etc. in traditional Chinese medicine pills.
  • the excipient may comprise the base part of a semi-solid formulation ointment, cream.
  • the excipients may include preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solubilizers, osmotic pressure regulators, colorants in liquid formulations.
  • the present application provides a method for detecting or measuring MASP-2, which may include using the isolated antigen-binding protein or the polypeptide.
  • the methods may include in vitro methods, ex vivo methods, methods for non-diagnostic or non-therapeutic purposes.
  • the methods may include methods of detecting the presence and/or amount of MASP-2 for non-diagnostic purposes, which may include the steps of:
  • the present application provides a MASP-2 kit, which may include the use of the isolated antigen-binding protein or the polypeptide.
  • the kit may further include instructions for use, which describe the method for detecting the presence and/or content of MASP-2.
  • the methods may include in vitro methods, ex vivo methods, methods for non-diagnostic or non-therapeutic purposes.
  • the present application provides a use of the isolated antigen-binding protein or the polypeptide in the preparation of a kit, and the kit can be used in a method for detecting the presence and/or content of MASP-2.
  • the methods may include in vitro methods, ex vivo methods, methods for non-diagnostic or non-therapeutic purposes.
  • the application provides the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the carrier, and the pharmaceutical composition for preventing, alleviating and/or treat a disease or condition.
  • the application provides a kind of said isolated antigen binding protein, said polypeptide, said immunoconjugate, said isolated nucleic acid molecule, said carrier, said cell and /or the use of the pharmaceutical composition in the preparation of medicaments for preventing, alleviating and/or treating diseases or conditions.
  • the present application provides a method for preventing and/or treating a disease or disorder, comprising administering the isolated antigen-binding protein, the isolated nucleic acid molecule, the The carrier, the cell, the pharmaceutical composition.
  • compositions, pharmaceutical combinations and methods described herein may be used in conjunction with other types of cancer therapy, such as chemotherapy, surgery, radiation, gene therapy, and the like.
  • the pharmaceutical compositions and methods described in this application can be used in other disease conditions that depend on immune responses, such as inflammation, immune diseases and infectious diseases.
  • the subject may include humans or non-human animals.
  • the non-human animal can be selected from the group consisting of monkeys, chickens, geese, cats, dogs, mice and rats.
  • non-human animals may also include any animal species other than humans, such as livestock animals, or rodents, or primates, or domestic animals, or poultry animals.
  • the human can be Caucasian, African, Asian, Semitic, or other ethnicity, or a hybrid of various ethnicities.
  • the human can be elderly, adult, adolescent, child or infant.
  • the effective amount in humans can be inferred from the effective amount in experimental animals.
  • Freireich et al. describe the correlation of doses in animals and humans (based on milligrams per square meter of body surface) (Freiheim et al., Cancer Chemother. Rep. 50, 219 (1966)).
  • Body surface area can be approximately determined from the patient's height and weight. See, eg, Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y., 537 (1970).
  • Human MASP-2 protein (O00187) includes six domains.
  • domain 4 CPP1
  • domain 5 CPP2
  • SP domain 6
  • the expressed protein exists in the form of inclusion body.
  • the inclusion body is renatured and then purified by the C-terminal His of the protein fragment.
  • the obtained recombinant human MASP-2 proteins are hMASP2-D456 (the amino acid sequence of which is shown in SEQ ID NO: 1) and hMASP2A-D456 (the amino acid sequence of which is shown in SEQ ID NO: 2).
  • the above-mentioned hMASP2A-D456 and PADRE were emulsified with complete Freund's adjuvant at a ratio of 1:1, and then injected intraperitoneally into 6- to 8-week-old female SD rats, and injected intraperitoneally and subcutaneously at multiple sites. 6-8 weeks-old female Balb/c mice, 50 ⁇ g hMASP2A-D456/mouse; after that, booster immunization was carried out at intervals of two to three weeks, and each rat was intraperitoneally injected with 50 ⁇ g antigen plus Freund’s incomplete adjuvant.
  • each mouse was intraperitoneally injected with 50 ⁇ g of antigen plus incomplete Freund's adjuvant. A total of 3 times of immunization, the tail blood was taken two weeks after the last immunization, and the serum anti-human MASP-2 antibody titer was determined. After the animals to be fused were immunized by intraperitoneal injection of 50 ⁇ g protein without Freund's adjuvant, spleen cells were collected on the third day.
  • hMASP2-D456 Dilute hMASP2-D456 to 1.0 ⁇ g/mL with carbonate buffer, add to a high-binding transparent polystyrene 96-well plate (Nunc), 100 ⁇ L/well, and coat at 4°C overnight. The next day the ELISA plate was washed twice with wash buffer (PBS+0.05% Tween 20 (sigma)) on an automatic plate washer. Add 300 ⁇ L of blocking buffer (PBS+0.05% Tween 20 (sigma)+1% BSA) to each well, and block at room temperature for 1 hour.
  • wash buffer PBS+0.05% Tween 20 (sigma)
  • BSA blocking buffer
  • Thermo Multiscan FC read the plate on Thermo Multiscan FC at 450 nm.
  • the hybridoma clone rats were screened: 50A6; mouse 47A1; hybridoma monoclonal antibodies were prepared by serum-free culture and conventional antibody purification methods for antibody function confirmation.
  • hMASP2-D456 Dilute hMASP2-D456 with carbonate buffer solution (CBS) to 1.0 ⁇ g/mL and 0.5 ⁇ g/mL, add to a high-binding transparent polystyrene 96-well plate (Nunc), 100 ⁇ L/well, and coat overnight at 4°C. The next day the ELISA plate was washed twice with wash buffer (PBS+0.05% Tween 20 (sigma)) on an automatic plate washer. Add 300 ⁇ L of blocking buffer (PBS+0.05% Tween 20 (sigma)+1% BSA) to each well, and block at room temperature for 1 hour. Then wash twice with washing buffer on an automatic plate washer, and dilute the monoclonal antibody to 8 gradients with blocking buffer.
  • CBS carbonate buffer solution
  • the 50A6 antibody can bind to human MASP-2; as shown in Figure 2, the 47A1 antibody can bind to human MASP-2, and the mouse anti-human MASP-2 monoclonal antibodies of the present application can all bind to human MASP-2.
  • 50A6 was selected as the candidate antibody of rat origin and humanized
  • 47A1 was selected as the candidate antibody of mouse origin and humanized.
  • Embodiment 4 Humanization of hybridoma monoclonal antibody
  • the antibody light and heavy chain variable regions are numbered by Chothia (see Chothia & Lesk, 1987) to define antibody CDR regions: CDRL1 (L24-L34, which means amino acids 24-34 of VL), CDRL2 (L50-L56), CDRL3 (L89-L97 ), CDRH1 (H26-H32, which means the 26th-32nd amino acid of VH), CDRH2 (H52-H56), CDRH3 (H95-H97), according to the sequence alignment and variable region structure information, the antibody light and heavy chains are variable Region amino acids were subjected to humanization mutations.
  • the light chain variable region sequence of 47A1 (its amino acid sequence is shown in SEQ ID NO: 31); the heavy chain variable region sequence of 47A1 (its amino acid sequence is shown in SEQ ID NO: 39).
  • Amino acid mutations were then carried out on the basis of the amino acid sequence of the variable region of the above-mentioned chimeric antibody to obtain the following humanized optimized amino acid sequence: the light chain variable region sequence of JYB1931A63 (the amino acid sequence is shown in SEQ ID NO: 21) , the heavy chain variable region sequence of JYB1931A63 (its amino acid sequence is shown in SEQ ID NO: 26).
  • the light chain variable region sequence of JYB1931A13 its amino acid sequence is shown in SEQ ID NO:47
  • the heavy chain variable region sequence of JYB1931A13 shows amino acid sequence.
  • ExpiCHO cells Cultivate ExpiCHO cells at a concentration of 8% carbon dioxide to a density of 6 ⁇ 10 6 /mL, and use ExpiFectamine transfection reagent to transfect 10 ⁇ g of antibody light and heavy chain expression plasmids into the cells; one day after transfection, take 150 ⁇ L and 4 mL of ExpiCHO Enhancer and ExpiCHO The auxiliary materials were added to the cultured cells, and the culture was continued for 9 days, and the supernatant was collected by centrifugation at 3500 rpm at 4°C.
  • the complement system lectin pathway (Svar Life Science AB, catalog number: AS 1327) kit is used to detect the blockage of the antibody on the complement signaling pathway in human serum.
  • Octet RED96e (Fortebio) was used to determine the affinity of candidate antibodies JYB1931A63 and JYB1931A13 to human hMASP2-D456 (KACTUS, lot number: 080203), monkey cMASP2-D456 (KACTUS, lot number: 030301) and mouse mMASP2-D456 (KACTUS, lot number: 030401) , the antigen and antibody were diluted with 1xPBST (1xPBS: Sanko, B548117-0500; 0.02% Tween 20: sigma-aldrich, P1379), the concentration of antigen used was 30 nM, and the concentration of antibody used was 5 ⁇ g/mL.
  • 1xPBST 1xPBS: Sanko, B548117-0500
  • Tween 20 sigma-aldrich
  • the sample was added to a 96-well plate (Greiner bio-one, 655209), and the system was 200 ⁇ L/well. Then set the software parameters, the plate temperature is set to 30°C, and the frequency of collecting standard kinetic signals is 5.0HZ. Next, pre-wet the AHC sensor (Fortébio, catalog number: 18-0015) with 1xPBST for 10 minutes, and then test it on the machine.
  • Each cycle includes the following steps: 1) immersion in buffer for 60s; 2) detection of non-specific binding of antigen to the sensor; 3) regeneration of 10mM glycine solution at pH 1.7; 4) immersion in buffer for 60s; 5) antibody immobilization on the sensor , the time is 60s; 6) the sensor is immersed in the buffer for 180s; 7) the combination of antigen and antibody, the time is 180s; 8) the dissociation of antigen and antibody, the time is 5 minutes; 9) sensor regeneration.
  • Complement system lectin pathway (Svar Life Science AB, catalog number: AS 1327) kit detects the complement signal pathway blocking of human serum and monkey serum by antibodies.
  • Samples were diluted with three diluents: 500nM, 10-fold dilution.
  • the experimental steps and data processing are the same as in Example 5.
  • the humanized antibody of the present application can block the lectin pathway of the complement system.
  • test product Dilute the test product with sample buffer to 1 mg/mL, then follow the instructions of the Protein Thermal Shift TM Starter Kit, take 13 ⁇ L of the test solution and add it to the PCR tube, add 5 ⁇ L Protein Thermal shift TM Buffer, add 2 ⁇ L 10 ⁇ staining solution, and make The reaction volume was 20 ⁇ L. After mixing, centrifuge at 12000 rpm for 5 min to remove air bubbles. Put the detection sample in the PCR instrument, perform sample analysis, and record the Tm value of the sample.
  • Humanized FcRn mice were used as experimental animals to study the pharmacokinetic indicators of two positive controls (Narsoplimab) and JYB1931A63 after a single intraperitoneal administration. All animal experimental protocols were reviewed and approved by IACUC.
  • hFcRn mice were purchased from Beijing Biocytogen, male, 6-8 weeks old, weighing 23-26g, raised in SPF grade animal room, fed with standard pellet feed, free to eat and drink, room temperature 18-24°C, relative humidity 40% ⁇ 50%, 12 hours a day and night.
  • a total of 16 experimental animals were randomly divided into four groups, four animals in each group, administered intraperitoneally once, with a dose of 10 mg/kg and a volume of 10 mL/kg.
  • the blood collection time points were before administration, 2h, 6h, 24h (day 1), day 2, day 3, day 4, day 7, day 10, day 14, day 21 after administration, Day 28, Day 35 and Day 42. Collect 60 ⁇ L/piece of whole blood through the orbit into EP tubes, let it stand at room temperature for 30 minutes, then centrifuge (2000g, 4°C, 5 minutes) to separate serum, and divide each sample into 2 parts (detection tube and backup tube), 10 ⁇ L/tube , stored at -80°C.
  • the pharmacokinetics of the four drugs at different time points were analyzed by Elisa indirect method.
  • Coating antigen is recombinant human MASP-2 protein, 1 ⁇ g/mL, 100 ⁇ L/well, 4°C, overnight. After washing the plate, use 200 ⁇ L/well blocking solution overnight at 4°C. Add serum samples, 50 ⁇ L/well, 37°C, 1h.
  • the detection antibody is mouse monoclonal antibody [H2] anti-human IgG F(ab)'2 (HRP) (abcam, ab87422, GR3246767-11) + Streptavidin-peroxidase (Sigma, lot: SLCB5784), 100 ⁇ l/well, 37°C , 0.5h.
  • TMB Chromogenic Solution (KPL, Cat. No.: 52-00-03) was developed, and the microplate reader (Molecular Devices, SpectraMax M3) read OD450.
  • the drug concentration was obtained according to the standard curve, and the PK Solver performed non-compartmental data processing to obtain the PK parameters.
  • the plasma concentration curve is shown in Figure 7.

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Abstract

一种抗MASP-2抗体及其用途，所述抗MASP-2抗体以约2E-09M或以下的K D值与人MASP-2蛋白特异性结合，所述抗MASP-2抗体以约2E-09M或以下的K D值与猴MASP-2蛋白特异性结合。一种包含所述抗MASP-2抗体的免疫缀合物、制备所述抗MASP-2抗体的方法以及抗MASP-2抗体的用途。

Description

抗MASP-2抗体及其用途 技术领域
本申请涉及生物医药领域，具体的涉及一种抗MASP-2抗体及其用途。
背景技术
免疫球蛋白A肾病(IgAN)(也称为Berger病)是一种系膜增生性肾小球肾炎(GN)，其特征是IgA在肾系膜内弥漫性沉积。目前对IgA肾病的治疗，KIDGO指南推荐使用ACEi/ARB降低尿蛋白，改善肾脏功能，同时，免疫荧光研究显示肾小球局部补体C3活化与预后不良有关，这暗示IgA肾病和补体系统活化有关，补体系统包含三条通路：1)经典途径(CP classical pathway)；2)旁路途径(AP alternative pathway)；3)凝集素途径(LP lectin pathway)。以前医学界普遍认为这个疾病预后好，不会对健康造成损害，但随着时间推移，对IgA肾病的深入了解，医生们发现，并不是所有IgA肾病患者预后都那么好，有相当一部分IgA肾病患者在发病后10-20年发展成尿毒症。如果不控制好进展速度快的IgA肾病，会给家庭和社会带来巨大的负担。但目前治疗IgA肾病的“武器”比较少，RAS阻断剂(沙坦和普利类)、激素虽然能控制IgA肾病患者的病情，但有相当一部分患者对这些药物的应答不佳。补体异常活化导致肾脏损伤是目前基础研究很重要的方向，而部分IgA肾病加重就是由于补体异常活化所致。
MASP-2是补体系统(complement system)中凝集素信号通路的效应酶(effector enzyme)，是阻止补体异常活化的理想靶点之一。
因此，亟需开发对MASP-2亲和力高、特异性强的新型抗MASP-2抗体。
发明内容
本申请提供了一种分离的抗原结合蛋白，其具有下述性质中的一种或多种：1)在Octet检测中，以约2E-09M或以下的KD值与人MASP-2蛋白特异性结合；2)在Octet检测中，以约2E-09M或以下的KD值与猴MASP-2蛋白特异性结合；以及3)能够特异性阻断人补体系统凝集素通路而不影响补体经典路径和旁路途径。
在某些实施方式中，所述分离的抗原结合蛋白包含HCDR3，所述HCDR3包含SEQ ID NO：19所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含HCDR2，所述HCDR2包含SEQ ID NO：17所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含HCDR1，所述HCDR1包含SEQ ID NO：15所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO：67所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO：13和SEQ ID NO：26所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包括重链可变区VH，所述VH包含所述HCDR1、HCDR2和HCDR3，所述HCDR3包含SEQ ID NO：19所示的氨基酸序列；所述HCDR2包含SEQ ID NO：17所示的氨基酸序列；以及所述HCDR1包含SEQ ID NO：15所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR1，所述H-FR1的C末端与所述HCDR1的N末端直接或间接地相连，且所述H-FR1包含SEQ ID NO：62所示的氨基酸序列。
在某些实施方式中，所述H-FR1包含SEQ ID NO:14和SEQ ID NO:27中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR2，所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO：63所示的氨基酸序列。
在某些实施方式中，所述H-FR2包含SEQ ID NO:16和SEQ ID NO:28中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR3，所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO：64所示的氨基酸序列。
在某些实施方式中，所述H-FR3包含SEQ ID NO:18和SEQ ID NO:29中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR4，所述H-FR4的N末端与所述HCDR3的C末端直接或间接地相连，且所述H-FR4包含SEQ ID NO：65所示的氨基酸序 列。
在某些实施方式中，所述H-FR4包含SEQ ID NO:20和SEQ ID NO:30中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR1，H-FR2，H-FR3和H-FR4，所述H-FR1包含SEQ ID NO：62所示的氨基酸序列；所述H-FR2包含SEQ ID NO：63所示的氨基酸序列；所述H-FR3包含SEQ ID NO：64所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO：65所示的氨基酸序列。
在某些实施方式中，所述H-FR1包含SEQ ID NO:14和SEQ ID NO:27中任一项所示的氨基酸序列；所述H-FR2包含SEQ ID NO:16和SEQ ID NO:28中任一项所示的氨基酸序列；所述H-FR3包含SEQ ID NO:18和SEQ ID NO:29中任一项所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO:20和SEQ ID NO:30中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述H-FR1、H-FR2、H-FR3和H-FR4包含选自下述任意一组的氨基酸序列：
a)H-FR1：SEQ ID NO:14，H-FR2：SEQ ID NO:16，H-FR3：SEQ ID NO:18和H-FR4：SEQ ID NO:20；
b)H-FR1：SEQ ID NO:27，H-FR2：SEQ ID NO:28，H-FR3：SEQ ID NO:29和H-FR4：SEQ ID NO:30。
在某些实施方式中，所述分离的抗原结合蛋白包含重链可变区VH，所述VH包含SEQ ID NO：67所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含重链可变区VH，所述VH包含SEQ ID NO:13和SEQ ID NO:26中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含LCDR3，所述LCDR3包含SEQ ID NO：11所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含LCDR2，所述LCDR2包含SEQ ID NO：9所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含LCDR1，所述LCDR1包含SEQ ID NO：7所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO：66所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO：5和21中任一项所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包含轻链可变区VL，所述VL包含所述LCDR1、LCDR2和LCDR3，所述LCDR3包含SEQ ID NO：11所示的氨基酸序列；所述LCDR2包含SEQ ID NO：9所示的氨基酸序列；以及所述LCDR1包含SEQ ID NO：7所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR1，所述L-FR1的C末端与所述LCDR1的N末端直接或间接地相连，且所述L-FR1包含SEQ ID NO：58所示的氨基酸序列。
在某些实施方式中，所述L-FR1包含SEQ ID NO:6和SEQ ID NO:22中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR2，所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO：59所示的氨基酸序列。
在某些实施方式中，所述L-FR2包含SEQ ID NO:8和SEQ ID NO:23中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR3，所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO：60所示的氨基酸序列。
在某些实施方式中，所述L-FR3包含SEQ ID NO:10和SEQ ID NO:24中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR4，所述L-FR4的N末端与所述LCDR3的C末端直接或间接地相连，且所述L-FR4包含SEQ ID NO：61所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR4包含SEQ ID NO:12和SEQ ID NO:25中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR1，L-FR2，L-FR3和L-FR4，所述L-FR1包含SEQ ID NO：58所示的氨基酸序列；所述L-FR2包含SEQ ID NO：59所示的氨基酸序列；所述L-FR3包含SEQ ID NO：60所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO：61所示的氨基酸序列。
在某些实施方式中，所述L-FR1包含SEQ ID NO:6和SEQ ID NO:22中任一项所示的氨基酸序列；所述L-FR2包含SEQ ID NO:8和SEQ ID NO:23中任一项所示的氨基酸序列；所述L-FR3包含SEQ ID NO:10和SEQ ID NO:24中任一项所示的氨基酸序列；以及所述L- FR4包含SEQ ID NO:12和SEQ ID NO:25中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR1、L-FR2、L-FR3和L-FR4包含选自下述任意一组的氨基酸序列：
a)L-FR1：SEQ ID NO:6，L-FR2：SEQ ID NO:8，L-FR3：SEQ ID NO:10和L-FR4：SEQ ID NO:12；
b)L-FR1：SEQ ID NO:22，L-FR2：SEQ ID NO:23，L-FR3：SEQ ID NO:24和L-FR4：SEQ ID NO:25。
在某些实施方式中，所述分离的抗原结合蛋白中所述VL包含SEQ ID NO：66所示的氨基酸序列。
在某些实施方式中，所述VL包含SEQ ID NO:5和SEQ ID NO:21中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述VH和VL包含选自下述任意一组的氨基酸序列：
a)VH：SEQ ID NO：13和VL：SEQ ID NO：5；
b)VH：SEQ ID NO：26和VL：SEQ ID NO：21。
在某些实施方式中，所述分离的抗原结合蛋白包含HCDR3，所述HCDR3包含SEQ ID NO：45所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含HCDR2，所述HCDR2包含SEQ ID NO：43所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含HCDR1，所述HCDR1包含SEQ ID NO：41所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO：77所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO:39和SEQ ID NO:52中任一项所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包括重链可变区VH，所述VH包含所述HCDR1、HCDR2和HCDR3，所述HCDR3包含SEQ ID NO：45所示的氨基酸序列；所述HCDR2包含SEQ ID NO：43所示的氨基酸序列；以及所述HCDR1包含SEQ ID NO：41所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR1，所述H-FR1的C末端与所述HCDR1的N末端直接或间接地相连，且所述H-FR1包含SEQ ID NO：72所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述H-FR1包含SEQ ID NO:40和SEQ ID NO:53中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR2，所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO：73所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述H-FR2包含SEQ ID NO:42和SEQ ID NO:54中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR3，所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO：74所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述H-FR3包含SEQ ID NO:44和SEQ ID NO:55中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR4，所述H-FR4的N末端与所述HCDR3的C末端直接或间接地相连，且所述H-FR4包含SEQ ID NO：75所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR4，所述H-FR4的N末端与所述HCDR3的C末端直接或间接地相连，且所述H-FR4包含SEQ ID NO:46和SEQ ID NO:30中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含H-FR1，H-FR2，H-FR3和H-FR4，所述H-FR1包含SEQ ID NO：72所示的氨基酸序列；所述H-FR2包含SEQ ID NO：73所示的氨基酸序列；所述H-FR3包含SEQ ID NO：74所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO：75所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述H-FR1包含SEQ ID NO:40和SEQ ID NO:53中任一项所示的氨基酸序列；所述H-FR2包含SEQ ID NO:42和SEQ ID NO:54中任一项所示的氨基酸序列；所述H-FR3包含SEQ ID NO:44和SEQ ID NO:55中任一项所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO:46和SEQ ID NO:30中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述H-FR1、H-FR2、H-FR3和H-FR4 包含选自下述任意一组的氨基酸序列：
a)H-FR1：SEQ ID NO:40，H-FR2：SEQ ID NO:42，H-FR3：SEQ ID NO:44和H-FR4：SEQ ID NO:46；
b)H-FR1：SEQ ID NO:53，H-FR2：SEQ ID NO:54，H-FR3：SEQ ID NO:55和H-FR4：SEQ ID NO:30。
在某些实施方式中，所述分离的抗原结合蛋白包含重链可变区VH，所述VH包含SEQ ID NO：77所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述VH包含SEQ ID NO:39和SEQ ID NO:52中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含LCDR3，所述LCDR3包含SEQ ID NO：37所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含LCDR2，所述LCDR2包含SEQ ID NO：35所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含LCDR1，所述LCDR1包含SEQ ID NO：33所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO：76所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包含SEQ ID NO:31和SEQ ID NO:47中任一项所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
在某些实施方式中，所述分离的抗原结合蛋白包含轻链可变区VL，所述VL包含所述LCDR1、LCDR2和LCDR3，所述LCDR3包含SEQ ID NO：37所示的氨基酸序列；所述LCDR2包含SEQ ID NO：35所示的氨基酸序列；以及所述LCDR1包含SEQ ID NO：33所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR1，所述L-FR1的C末端与所述LCDR1的N末端直接或间接地相连，且所述L-FR1包含SEQ ID NO：68所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR1包含SEQ ID NO:32和SEQ ID NO:48中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR2，所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO：69所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR2包含SEQ ID NO:34和SEQ ID NO:49中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR3，所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO：70所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR3包含SEQ ID NO:36和SEQ ID NO:50中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR4，所述L-FR4的N末端与所述LCDR3的C末端直接或间接地相连，且所述L-FR4包含SEQ ID NO：71所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR4包含SEQ ID NO:38和SEQ ID NO:51中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含L-FR1，L-FR2，L-FR3和L-FR4，所述L-FR1包含SEQ ID NO：68所示的氨基酸序列；所述L-FR2包含SEQ ID NO：69所示的氨基酸序列；所述L-FR3包含SEQ ID NO：70所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO：71所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR1包含SEQ ID NO:32和SEQ ID NO:48中任一项所示的氨基酸序列；所述L-FR2包含SEQ ID NO:34和SEQ ID NO:49中任一项所示的氨基酸序列；所述L-FR3包含SEQ ID NO:36和SEQ ID NO:50中任一项所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO:38和SEQ ID NO:51中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述L-FR1、L-FR2、L-FR3和L-FR4包含选自下述任意一组的氨基酸序列：
a)L-FR1：SEQ ID NO:32，L-FR2：SEQ ID NO:34，L-FR3：SEQ ID NO:36和L-FR4：SEQ ID NO:38；
b)L-FR1：SEQ ID NO:48，L-FR2：SEQ ID NO:49，L-FR3：SEQ ID NO:50和L-FR4：SEQ ID NO:51。
在某些实施方式中，所述分离的抗原结合蛋白包括VL，所述VL包含SEQ ID NO：76所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白中所述VL包含SEQ ID NO：31和47中任一项所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包括VH和VL，所述VH和VL包含选自下述任意一组的氨基酸序列：
a)VH：SEQ ID NO:39和VL：SEQ ID NO:31；
b)VH：SEQ ID NO:52和VL：SEQ ID NO:47。
在某些实施方式中，所述分离的抗原结合蛋白包含重链恒定区，且所述重链恒定区包括源自IgG的恒定区或源自IgY的恒定区。
在某些实施方式中，所述分离的抗体结合蛋白中所述重链恒定区包括源自IgG的恒定区。
在某些实施方式中，所述分离的抗原结合蛋白中所述重链恒定区包括源自IgG1、IgG2、IgG3或IgG4的恒定区。
在某些实施方式中，所述分离的抗原结合蛋白中所述重链恒定区包含SEQ ID NO：56所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包含轻链恒定区，且所述轻链恒定区包括源自Igκ的恒定区或源自Igλ的恒定区。
在某些实施方式中，所述分离的抗原结合蛋白中所述轻链恒定区包括源自人Igκ的恒定区。
在某些实施方式中，所述分离的抗原结合蛋白中所述轻链恒定区包含SEQ ID NO：57所示的氨基酸序列。
在某些实施方式中，所述分离的抗原结合蛋白包括抗体或其抗原结合片段。
在某些实施方式中，所述分离的抗原结合蛋白中所述抗原结合片段选自下组：Fab，Fab’，F(ab)2，Fv片段，F(ab’)2，scFv，di-scFv，VHH和/或dAb。
在某些实施方式中，所述分离的抗原结合蛋白中所述抗体选自下组：单克隆抗体、单链抗体、嵌合抗体、人源化抗体和全人源抗体。
另一方面，本申请提供了一种或多种多肽，其包含所述分离的抗原结合蛋白。
另一方面，本申请提供了一种或多种免疫缀合物，其包含所述分离的抗原结合蛋白或所述的多肽。
另一方面，本申请提供了一种或多种分离的核酸分子，其编码所述分离的抗原结合蛋白，或者所述的多肽。
另一方面，本申请提供了一种或多种载体，其包含所述分离的核酸分子。
另一方面，本申请提供了一种或多种细胞，其包含所述分离的抗原结合蛋白，所述的多 肽，所述的免疫缀合物，所述分离的核酸分子和/或所述的载体。
另一方面，本申请提供了一种制备所述分离的抗原结合蛋白或所述的多肽的方法，所述方法包括再使得所述分离的抗原结合蛋白或所述的多肽表达的条件下，培养所述的细胞。
另一方面，本申请提供了一种或多种药物组合物，其包含所述分离的抗原结合蛋白，所述的多肽，所述的免疫缀合物，所述分离的核酸分子，所述的载体，所述的细胞，和/或药学上可接受的佐剂和/或赋形剂。
另一方面，本申请提供了一种用于检测或测定MASP-2的方法，所述方法包括使用所述分离的抗原结合蛋白或所述的多肽。
另一方面，本申请提供了一种MASP-2的检测试剂盒，其包含所述分离的抗原结合蛋白或所述的多肽。
另一方面，本申请提供了一种分离的抗原结合蛋白或所述的多肽在制备试剂盒中的用途，所述试剂盒用于检测MASP-2的存在和/或含量。
另一方面，本申请提供了一种所述分离的抗原结合蛋白，所述的多肽，所述的免疫缀合物，所述分离的核酸分子，所述的载体，所述的细胞和/或所述药物组合物在制备预防和/或治疗疾病或病症的药物中的用途。
另一方面，本申请提供了所述分离的抗原结合蛋白，所述的多肽，所述的免疫缀合物，所述分离的核酸分子，所述的载体，所述的细胞和/或所述药物组合物，其用于预防、缓解和/或治疗疾病或病症。
另一方面，本申请提供了一种预防和/或治疗疾病或病症的方法，其包括向有需要的受试者施用有效量的所述分离的抗原结合蛋白，所述的多肽，所述的免疫缀合物，所述分离的核酸分子，所述的载体，和/或所述的细胞。
本领域技术人员能够从下文的详细描述中容易地洞察到本申请的其它方面和优势。下文的详细描述中仅显示和描述了本申请的示例性实施方式。如本领域技术人员将认识到的，本申请的内容使得本领域技术人员能够对所公开的具体实施方式进行改动而不脱离本申请所涉及发明的精神和范围。相应地，本申请的附图和说明书中的描述仅仅是示例性的，而非为限制性的。
附图说明
本申请所涉及的发明的具体特征如所附权利要求书所显示。通过参考下文中详细描述的 示例性实施方式和附图能够更好地理解本申请所涉及发明的特点和优势。对附图简要说明如下：
图1显示的是本申请所述示例性的抗MASP-2抗体50A6与人MASP-2的结合。
图2显示的是本申请所述示例性的抗MASP-2抗体47A1与人MASP-2的结合。
图3A显示的是本申请所述示例性的抗MASP-2抗体50A6与食蟹猴MASP-2的结合。
图3B显示的是本申请所述示例性的抗MASP-2抗体50A6与鼠MASP-2的结合。
图4A显示的是本申请所述示例性的抗MASP-2抗体47A1与食蟹猴MASP-2的结合。
图4B显示的是本申请所述示例性的抗MASP-2抗体47A1与鼠MASP-2的结合。
图5显示的是本申请所述示例性的抗MASP-2抗体阻断补体系统凝集素通路的结果。
图6显示的是本申请所述示例性的人源化抗MASP-2抗体阻断补体系统凝集素通路的结果。
图7显示的是本申请所述示例性的抗MASP-2抗体在人源化FcRn小鼠模型中药代动力学(PK)的结果。
具体实施方式
以下由特定的具体实施例说明本申请发明的实施方式，熟悉此技术的人士可由本说明书所公开的内容容易地了解本申请发明的其他优点及效果。
术语定义
在本申请中，术语“分离的”通常指从天然状态下经人工手段获得的。如果自然界中出现某一种“分离”的物质或成分，那么可能是其所处的天然环境发生了改变，或从天然环境下分离出该物质，或二者情况均有发生。例如，某一活体动物体内天然存在某种未被分离的多聚核苷酸或多肽，而从这种天然状态下分离出来的高纯度的相同的多聚核苷酸或多肽即称之为分离的。术语“分离的”不排除混有人工或合成的物质，也不排除存在不影响物质活性的其它不纯物质。
在本申请中，术语“抗原结合蛋白”通常是指一种能够特异性识别和/或中和特定抗原的多肽分子。例如，在本申请中，术语“抗原结合蛋白”可包括“抗体”或“抗原结合片段”。例如，所述抗体可包含通过二硫键相互连接的至少两条重(H)链和两条轻(L)链组成的免疫球蛋白，并且可包括任何包含其抗原结合部分的分子。术语“抗体”可包括单克隆抗体、抗体片段或抗 体衍生物，包括但不限于鼠源抗体、人抗体(全人源抗体)、人源化抗体、嵌合抗体、单链抗体(例如，scFv)，以及与抗原结合的抗体片段(例如，Fab、Fab’，VHH和(Fab)2片段)。术语“抗体”还可包括抗体的所有重组体形式，例如在原核细胞中表达的抗体、未糖基化的抗体以及本文所述的任何与抗原结合的抗体片段及其衍生物。每条重链可由重链可变区(VH)和重链恒定区构成。每条轻链可由轻链可变区(VL)和轻链恒定区构成。VH和VL区可进一步被区分为称为互补决定区(CDR)的高变区，它们散布在称为构架区(FR)的更保守的区域中。每个VH和VL可由三个CDR和四个FR区构成，它们从氨基端至羧基端可按以下顺序排列：FR1、CDR1、FR2、CDR2、FR3、CDR3和FR4。重链和轻链的可变区含有与抗原(例如，人MASP-2)相互作用的结合结构域。抗体的恒定区可介导该免疫球蛋白与宿主组织或因子的结合，所述宿主组织或因子包括免疫系统的多种细胞(例如，效应细胞)和经典补体系统的第一成分(Clq)。所述CDRs的确切边界已根据不同系统不同地限定。由Kabat(Kabat等人，Sequences of Proteins of Immunological Interest(National Institutes of Health，Bethesda，Md.(1987)和(1991))描述的系统，不仅提供了可应用于抗原结合片段的任何可变区的明确残基编号系统，还提供了限定CDRs的精确残基边界。这些CDRs可以被称为Kabat CDRs。Chothia和同事(Chothia和Lesk，J.Mol.Biol.196：901-917(1987)以及Chothia等人，Nature 342：877-883(1989))发现尽管在氨基酸序列水平上具有大的多样性，但是Kabat CDRs内的某些亚部分采取几乎相同的肽主链构象。这些亚部分命名为L1、L2和L3或H1、H2和H3，其中“L”和“H”分别指轻链和重链区域。这些区域可以被称为Chothia CDRs，所述Chothia CDRs具有与Kabat CDRs重叠的边界。与Kabat CDRs重叠的限定CDRs的其他边界已由Padlan(FASEB J.9：133-139(1995))和MacCallum(J Mol Biol 262(5)：732-45(1996))描述。另外，其他的CDR边界定义可能不严格地遵循上述系统之一，但仍将与Kabat CDRs重叠，尽管按照特定残基或残基组或甚至整个CDRs并不显著影响抗原结合的预测或实验发现，它们可以缩短或加长。在本申请中，所述CDR可通过使用的是Chothia编号系统定义。
在本申请中，术语“抗原结合片段”通常是指抗体中发挥特异性结合抗原功能的一个或多个片段。抗体的抗原结合功能可通过抗体的全长片段来实现。抗体的抗原结合功能也可通过以下来实现：包括Fv、ScFv、dsFv、Fab、Fab’或F(ab’)2的片段的重链，或者，包括Fv、scFv、dsFv、Fab、Fab’或F(ab’)2的片段的轻链。(1)Fab片段，通常为由VL、VH、CL和CH结构域组成的一价片段；(2)F(ab’)2片段，包含通过铰链区处的二硫键连接的两个Fab片段的二价片段；(3)由VH和CH结构域组成的Fd片段；(4)由抗体单臂的VL和VH 结构域组成的Fv片段；(5)由VH结构域组成的dAb片段(Ward等，(1989)Nature 341：544-546)；(6)分离的互补决定区(CDR)和(7)可任选地通过接头连接的两个或以上分离的CDR的组合。例如，还可包括由VL和VH配对形成的一价单链分子Fv(scFv)(参见Bird等(1988)Science 242：423-426；以及Huston等(1988)Proc.Natl.Acad.Sci.85：5879-5883)。例如，还可包括缺失抗体轻链而只有重链可变区的一类抗体VHH(例如，可参见康晓圳等，生物工程学报,2018,34(12):1974-1984)。所述“抗原结合部分”还可包括免疫球蛋白融合蛋白，所述融合蛋白包含选自以下的结合结构域：(1)与免疫球蛋白铰链区多肽融合的结合结构域多肽；(2)与铰链区融合的免疫球蛋白重链CH2恒定区；和(3)与CH2恒定区融合的免疫球蛋白重链CH3恒定区。
在本申请中，术语“单克隆抗体”通常是指一群基本同源的抗体，即包含该群的各个抗体除了可能的以微量存在的天然发生的突变之外是相同的。单克隆抗体是高度特异性的，直接针对单个抗原性位点。例如，所述单克隆抗体可以通过杂交瘤技术制备或者通过使用重组DNA方法在细菌、真核动物或植物细胞中产生。单克隆抗体也可以得自噬菌体抗体文库，使用例如Clackson etal.,Nature,352:624-628(1991)和Marks et al.,Mol.Biol.，222:581-597(1991)所述的技术进行。
在本申请中，术语“嵌合抗体”通常是指这样的抗体，其中每个重链或轻链氨基酸序列的一部分与来自特定物种的抗体中相应氨基酸序列同源，或者属于特定的类别，而该链的其余区段则与另一物种中的相应序列同源。例如，轻链和重链的可变区均来自一个动物物种(如小鼠、大鼠等)的抗体的可变区，而恒定部分则与来自另一物种(如人)的抗体序列同源。例如，为获得嵌合抗体，可利用非人源的B细胞或杂交瘤细胞产生可变区，而与其组合的恒定区则来自人。所述可变区具有易于制备的优点，并且其特异性不受与其组合的恒定区的来源的影响。同时，由于嵌合抗体的恒定区可来源于人类，因此嵌合在注射时抗体引发免疫应答的可能性会低于使用恒定区为非人来源的抗体。
在本申请中，术语“人源化抗体”通常是指一种嵌合抗体，其含有较少的来自非人免疫球蛋白的序列，从而降低异种抗体引入到人类中时的免疫原性，同时保持抗体的完全抗原结合亲和力和特异性。例如，可以使用CDR移植(Jones et al.,Nature 321:522(1986))及其变体；包括“重塑”(reshaping)，(Verhoeyen,et al.,1988 Science 239:1534-1536；Riechmann,et al.,1988 Nature 332:323-337；Tempest,et al.,Bio/Technol 1991 9:266-271)，“高度加成”(hyperchimerization)，(Queen,et al.,1989 Proc Natl Acad Sci USA 86:10029-10033；Co,et al., 1991 Proc Natl Acad Sci USA 88:2869-2873；Co,et al.,1992 J Immunol 148:1149-1154)和“贴面”(veneering)，(Mark,et al.,“Derivation of therapeutically active humanized and veneered anti-CD18 antibodies.”In:Metcalf B W,Dalton B J,eds.Cellular adhesion:molecular definition to therapeutic potential.New York:Plenum Press,1994:291-312)、表面重建(美国专利US5639641)等技术手段，对非人源的结合域进行人源化。如果其他区域，例如铰链区和恒定区结构域也源自非人来源，则这些区域也可以被人源化。
在本申请中，术语“鼠源抗体”通常是指可变区框架和CDR区得自小鼠种系免疫球蛋白序列的抗体。此外，如果抗体包含恒定区，其也得自小鼠种系免疫球蛋白序列。本申请的鼠源抗体可以包含不由小鼠种系免疫球蛋白序列编码的氨基酸残基，例如可以包括通过体外随机突变或点突变或通过体内体细胞突变而导入的突变。
在本申请中，术语“种系(Germline)序列”通常是指未重排的免疫球蛋白DNA序列的序列。
在本申请中，术语“在……之间”通常是指某种氨基酸片段的C端与第一氨基酸片段的N端直接或间接连接，并且其N端与第二氨基酸片段的C端直接或间接连接。在轻链中，例如，所述L-FR2的N末端与所述LCDR1的C末端直接或间接相连，且所述L-FR2的C末端与所述LCDR2的N末端直接或间接相连。又例如，所述L-FR3的N末端与所述LCDR2的C末端直接或间接相连，且所述L-FR3的C末端与所述LCDR3的N末端直接或间接相连。在重链中，例如，所述H-FR2的N末端与所述HCDR1的C末端直接或间接相连，且所述H-FR2的C末端与所述HCDR2的N末端直接或间接相连。又例如，所述H-FR3的N末端与所述HCDR2的C末端直接或间接相连，且所述H-FR3的C末端与所述HCDR3的N末端直接或间接相连。
在本申请中，术语“MASP-2蛋白”，“MASP-2”或“MASP-2抗原”可以互换使用，并且包括MASP-2的任何功能活性片段、变体和同源物，其由细胞天然表达或在用MASP-2基因转染的细胞上表达。在本申请中，MASP-2可以为人MASP-2，其在UniProt/Swiss-Prot中的登录号为O00187。例如，MASP-2可以为人MASP-2的功能活性片段。在本申请中，MASP-2可以为猴MASP-2或其功能活性片段。例如，所述“功能活性片段”可以包括保留至少一种天然存在的蛋白质的内源功能(例如，与本申请所述的抗原结合蛋白结合)的片段。例如，所述“功能活性片段”可以包括与本申请的抗原结合蛋白结合的结构域。在本申请中，MASP-2是补体系统凝聚素通路中一个关键调节因子。血浆中甘露聚糖结合凝集素(mannan-binding lectin， MBL)或纤维胶凝蛋白(ficolin，FCN)直接识别多种病原微生物表面的甘露糖、N-乙酰甘露糖、N-乙酰葡萄糖氨、岩藻糖等为末端糖基的糖结构。MBL-MASP复合物与病原体表面糖结构结合，使MASP-1、MASP-2被独立地激活。活化的MASP-2发挥其SP活性，裂解C4，所产生的C4b片段共价结合于病原体表面，通过与C2相互作用，使后者也被MASP-2裂解，形成C3转化酶C4b2a，继之活化补体经典途径；活化的MASP1能直接裂解C3产生C3b，在蛋白因子D和蛋白因子P的作用下，形成C3转化酶C3bBb或C3bBbP，并产生C5转化酶C3bBb3b，激活补体旁路途径。
在本申请中，术语“IgA肾病”通常是指肾小球系膜区的原发性肾小球病。例如，所述“IgA肾病”可以包括以IgA或IgA沉积为主，伴或不伴有其他免疫球蛋白在肾小球系膜区沉积的原发性肾小球病。例如，所述“IgA肾病”的病变类型可包括局灶节段性病变、毛细血管内增生性病变、系膜增生性病变、新月体病变及硬化性病变等。其临床表现为反复发作性肉眼血尿或镜下血尿，可伴有不同程度蛋白尿，部分患者可以出现严重高血压或者肾功能不全。
除了本文提到的特定蛋白质和核苷酸之外，本申请还可包括其功能活性片段、衍生物、类似物、同源物及其片段。
术语“功能活性片段”指与天然存在序列具有基本上同一的氨基酸序列或由基本上同一的核苷酸序列编码并能够具有天然存在序列的一种或多种活性的多肽。在本申请的上下文中，任何给定序列的功能活性片段是指其中残基的特定序列(无论是氨基酸或核苷酸残基)已经经过修饰而使得所述多肽或多核苷酸基本上保留至少一种内源功能的序列。可以通过天然存在的蛋白质和/或多核苷酸中存在的至少一个氨基酸残基和/或核苷酸残基的添加、缺失、取代、修饰、替换和/或变异来获得编码功能活性片段的序列，只要保持原来的功能活性即可。
在本申请中，术语“衍生物”通常是指本申请的多肽或多核苷酸而言包括自/对序列的一个(或多个)氨基酸残基的任何取代、变异、修饰、替换、缺失和/或添加，只要所得的多肽或多核苷酸基本上保留其至少一种内源功能。
在本申请中，术语“类似物”通常对多肽或多核苷酸而言，包括多肽或多核苷酸的任何模拟物，即拥有该模拟物模拟的多肽或多核苷酸的至少一种内源功能的化学化合物。
通常，可以进行氨基酸取代，例如至少1个(例如，1、2、3、4、5、6、7、8、9、10或20个以上)氨基酸取代，只要经修饰的序列基本上保持需要的活性或能力。氨基酸取代可包括使用非天然存在的类似物。
在本申请中，术语“同源物”通常是指与天然存在序列具有一定同源性的氨基酸序列或核 苷酸序列。术语“同源性”可以等同于序列“同一性”。同源序列可以包括可以与主题序列是至少80％、85％、90％、99.1％、99.2％、99.3％、99.4％、99.5％、99.6％、99.7％、99.8％或99.9％相同的氨基酸序列。通常，同源物将包含与主题氨基酸序列相同的活性位点等。同源性可以根据相似性(即具有相似化学性质/功能的氨基酸残基)来考虑，也可以在序列同一性方面表达同源性。在本申请中，提及的氨基酸序列或核苷酸序列的SEQ ID NO中的任一项具有百分比同一性的序列是指在所提及的SEQ ID NO的整个长度上具有所述百分比同一性的序列。为了确定序列同一性，可进行序列比对，其可通过本领域技术人员了解的各种方式进行，例如，使用BLAST、BLAST-2、ALIGN、NEEDLE或Megalign(DNASTAR)软件等。本领域技术人员能够确定用于比对的适当参数，包括在所比较的全长序列中实现最优比对所需要的任何算法。
用于本申请的蛋白质或多肽也可以具有氨基酸残基的缺失、插入或取代，所述氨基酸残基产生沉默的变化并导致功能上等同的蛋白质。可以根据残基的极性、电荷、溶解性、疏水性、亲水性和/或两性性质的相似性进行有意的氨基酸取代，只要保留内源性功能即可。例如，带负电荷的氨基酸包括天冬氨酸和谷氨酸；带正电荷的氨基酸包括赖氨酸和精氨酸；并且含具有相似亲水性值的不带电极性头基的氨基酸包括天冬酰胺、谷氨酰胺、丝氨酸、苏氨酸和酪氨酸。
在本申请中，术语“免疫缀合物”通常是指所述其他治疗剂与所述分离的抗原结合蛋白缀合(例如，通过连接分子共价相连)而形成的缀合物，该缀合物可以通过所述分离的抗原结合蛋白与靶细胞上的抗原的特异性结合，将所述其他治疗剂递送至靶细胞。此外，所述抗原也可以由所述靶细胞分泌，并位于所述靶细胞外的间隙。
在本申请中，术语“受试者”通常指人类或非人类动物，包括但不限于猫、狗、马、猪、奶牛、羊、兔、小鼠、大鼠或猴。
在本申请中，术语“核酸分子”通常是指从其天然环境中分离的或人工合成的任何长度的分离形式的核苷酸、脱氧核糖核苷酸或核糖核苷酸或其类似物。
在本申请中，术语“载体”通常是指一种核酸分子，该种核酸分子能够转运与其连接的另一核酸。所述载体可将插入的核酸分子转移到细胞中和/或细胞之间。所述载体可包括主要用于将DNA或RNA插入细胞中的载体、主要用于复制DNA或RNA的载体，以及主要用于DNA或RNA的转录和/或翻译的表达的载体。所述载体可以是当引入合适的细胞时能够转录并翻译成多肽的多核苷酸。通常，通过培养包含所述载体的合适的细胞，所述载体可以产生 期望的表达产物。在本申请中，所述载体可包含慢病毒载体。
在本申请中，术语“细胞”通常是指可以或已经含有包括本申请所述的核酸分子的质粒或载体，或者能够表达本申请所述的多肽或本申请所述的抗原结合蛋白的个体细胞，细胞系或细胞培养物。所述细胞可以包括单个细胞的子代。由于天然的，意外的或故意的突变，子代细胞与原始亲本细胞在形态上或在基因组上可能不一定完全相同，但能够表达本申请所述的多肽或抗原结合蛋白即可。所述细胞可以通过使用本申请所述的载体体外转染细胞而得到。所述细胞可以是原核细胞(例如大肠杆菌)，也可以是真核细胞(例如酵母细胞，例如COS细胞，中国仓鼠卵巢(CHO)细胞，HeLa细胞，HEK293细胞，COS-1细胞，NS0细胞或骨髓瘤细胞)。在一些实施方式中，所述细胞可以是免疫细胞。例如，所述免疫细胞可以选自下组：T细胞、B细胞、天然杀伤细胞(NK细胞)、巨噬细胞、NKT细胞、单核细胞、树突状细胞、粒细胞、淋巴细胞、白细胞和/或外周血单个核细胞。
在本申请中，术语“治疗”通常是指：(i)预防可能易患疾病、病症和/或病状、但尚未诊断出患病的患者出现该疾病、病症或病状；(ii)抑制该疾病、病症或病状，亦即遏制其发展；以及(iii)缓解该疾病、病症或病状，亦即使得该疾病、病症和/或病状和/或与该疾病、病症和/或病状相关联的症状消退。
在本申请中，术语“多肽”、“肽”、“蛋白”和“蛋白质”可互换地使用，通常是指具有任何长度的氨基酸的聚合物。该聚合物可以是直链或支链的，它可以包含修饰的氨基酸，并且可以被非氨基酸中断。这些术语还涵盖已经被修饰的氨基酸聚合物。这些修饰可以包含：二硫键形成、糖基化、脂化(lipidation)、乙酰化、磷酸化、或任何其他操纵(如与标记组分结合)。术语“氨基酸”包括天然的和/或非天然的或者合成的氨基酸，包括甘氨酸以及D和L旋光异构体、以及氨基酸类似物和肽模拟物。
在本申请中，术语“多核苷酸”、“核苷酸”、“核苷酸序列”、“核酸”和“寡核苷酸”可互换地使用，通常是指具有任何长度的核苷酸的聚合形式，如脱氧核糖核苷酸或核糖核苷酸、或其类似物。多核苷酸可具有任何三维结构，并且可以执行已知或未知的任何功能。以下是多核苷酸的非限制性实例：基因或基因片段的编码区或非编码区、根据连接分析定义的多个座位(一个座位)、外显子、内含子、信使RNA(mRNA)、转运RNA、核糖体RNA、短干扰RNA(siRNA)、短发夹RNA(shRNA)、micro-RNA(miRNA)、核酶、cDNA、重组多核苷酸、分支多核苷酸、质粒、载体、任何序列的分离的DNA、任何序列的分离的RNA、核酸探针、和引物。多核苷酸可以包含一个或多个经修饰的核苷酸，如甲基化的核苷酸和核苷酸类似物。 如果存在，可以在聚合物组装之前或之后进行核苷酸结构的修饰。核苷酸的序列可以被非核苷酸组分中断。多核苷酸可以在聚合后，如通过与标记的组分缀合来进一步修饰。
在本申请中，术语“K D”(同样地，“K D”或“K D”)通常指“亲和常数”或“平衡解离常数”，并指在滴定测量中在平衡时、或者通过将解离速率常数(kd)除以结合速率常数(ka)所获得的值。使用结合速率常数(ka)、解离速率常数(kd)和平衡解离常数(K D)表示结合蛋白(例如本申请所述的分离的抗原结合蛋白)对抗原(例如，MASP-2蛋白)的结合亲和力。确定结合和解离速率常数的方法为本领域熟知。使用基于荧光的技术提供了高灵敏度以及在生理缓冲液中在平衡时检查样品的能力。例如，可以通过Biacore(生物分子相互作用分析)测定(例如，可以从BIAcoreInternationalAB，aGEHealthcarecompany，Uppsala，瑞典获得的仪器)所述K D值，也可以使用其他实验途径和仪器例如Octet检测。另外，也可以使用可以从SapidyneInstruments(Boise，Idaho)获得的KinExA(动态排阻测定(KineticExclusionAssay))测定所述K D值，或者使用表面等离子共振仪(SPR)测定所述K D值。例如，也可以通过胺偶联试剂盒测定所述K D值。
在本申请中，术语“和/或”应理解为意指可选项中的任一项或可选项的两项。
在本申请中，术语“包含”通常是指包括明确指定的特征，但不排除其他要素。在某些情形中，“包含”也涵盖仅包括指定的组分的情况。例如，包含也表示为也表示“由……组成”的含义。
在本申请中，术语“约”通常是指在指定数值以上或以下0.5％-10％的范围内变动，例如在指定数值以上或以下0.5％、1％、1.5％、2％、2.5％、3％、3.5％、4％、4.5％、5％、5.5％、6％、6.5％、7％、7.5％、8％、8.5％、9％、9.5％、或10％的范围内变动。
在本申请中，术语“包括”通常是指包含、总括、含有或包涵的含义。在某些情况下，也表示“为”、“由……组成”的含义。
发明详述
本申请所述分离的抗原结合蛋白
一方面，本申请提供一种分离的抗原结合蛋白，其在Octet检测中，可以以约2E-09M或以下的K D值(例如，所述K D不高于约2E-09M、不高于约1.5E-09M、不高于约1E-09M、不高于约9E-10M、不高于约8E-10M、不高于约7E-10M、不高于约6E-10M、不高于约5E-10M、不高于约2E-10M、不高于约1E-10M、不高于约5E-11M、不高于约1E-11M或不高于5E-12M 或以下)与人MASP-2蛋白特异性结合。
一方面，本申请提供一种分离的抗原结合蛋白，其在Octet检测中，可以以约2E-09M或以下的K D值(例如，所述K D不高于约2E-09M、不高于约1.5E-09M、不高于约1E-09M、不高于约9E-10M、不高于约8E-10M、不高于约7E-10M、不高于约6E-10M、不高于约5E-10M、不高于约2E-10M、不高于约1E-10M、不高于约5E-11M、不高于约1E-11M或不高于5E-12M或以下)与猴MASP-2蛋白特异性结合。
一方面，本申请提供一种分离的抗原结合蛋白，其可以包含抗体重链可变区VH中的至少一个CDR，所述VH可以包含SEQ ID NO：67或SEQ ID NO：77所示的氨基酸序列。
例如，所述VH可以包含SEQ ID NO:13、26、39和52中任一项所示的氨基酸序列。在本申请中，所述分离的抗原结合蛋白的HCDR可以通过任何形式划分，只要VH与SEQ ID NO:13、26、39和52中任一项所示的氨基酸序列相同，以任何形式划分得到的HCDR都可落入本申请的保护范围内。
抗体的CDR又称互补决定区，是可变区的一部分。该区域的氨基酸残基可以与抗原或抗原表位接触。抗体CDR可以通过多种编码系统来确定，如CCG、Kabat、Chothia、IMGT、AbM、综合考虑Kabat/Chothia等。这些编码系统为本领域内已知，具体可参见，例如，http://www.bioinf.org.uk/abs/index.html#kabatnum。本领域技术人员可以根据抗体的序列和结构，用不同的编码系统确定出CDR区。使用不同的编码系统，CDR区可能存在差别。在本申请中，所述CDR涵盖根据任何CDR划分方式划分得到的CDR序列；也涵盖其变体，所述变体包括所述CDR的氨基酸序列经过取代、缺失和/或添加一个或多个氨基酸。例如1-30个、1-20个或1-10个，又例如1个、2个、3个、4个、5个、6个、7个、8个或9个氨基酸取代、缺失和/或插入；也涵盖其同源物，所述同源物可以为与所述CDR的氨基酸序列具有至少约85％(例如，具有至少约85％、约90％、约91％、约92％、约93％、约94％、约95％、约96％、约97％、约98％、约99％或更高的)序列同源性的氨基酸序列。在某些实施方式中，本申请所述分离的抗原结合蛋白通过Chothia编码系统定义。
在本申请中，所述抗原结合蛋白可包含重链可变区VH，所述VH可包含HCDR1、HCDR2和HCDR3中的至少一个、两个或三个。
在本申请中，所述抗原结合蛋白的HCDR3可包含SEQ ID NO：19所示的氨基酸序列。例如，所述抗原结合蛋白的HCDR3序列可根据Chothia编码系统定义。
在本申请中，所述抗原结合蛋白的HCDR2可包含SEQ ID NO：17所示的氨基酸序列。 例如，所述抗原结合蛋白的HCDR2序列可根据Chothia编码系统定义。
在本申请中，所述抗原结合蛋白的HCDR1可包含SEQ ID NO：15所示的氨基酸序列。例如，所述抗原结合蛋白的HCDR1序列可根据Chothia编码系统定义。
例如，所述抗原结合蛋白的HCDR1可包含SEQ ID NO：15所示的氨基酸序列；所述HCDR2可包含SEQ ID NO：17所示的氨基酸序列；且所述HCDR3可包含SEQ ID NO：19所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6、JYB1931A63或与其具有相同HCDR3(例如，与其具有相同的HCDR1-3)的抗原结合片段。
例如，所述抗原结合蛋白的VH可包含框架区H-FR1，H-FR2，H-FR3和H-FR4。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：62所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR1与SEQ ID NO：62所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 16和X 19。
EVQLVESGGGLVQPG X 16SL X 19LSCAAS(SEQ ID NO：62)，其中，X 16可以是G或R，X 19可以是R或S。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO:14和SEQ ID NO:27中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR2可包含SEQ ID NO：63所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR2与SEQ ID NO：63所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 8和X 10。
NMAWVRQ X 8P X 10KGLEWVATI(SEQ ID NO：63)，其中，X 8可以是A或T，X 10可以是G或K。
在本申请中，所述抗原结合蛋白的H-FR2可包含SEQ ID NO:16和SEQ ID NO:28中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR3可包含SEQ ID NO：64所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR3与SEQ ID NO：64所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 16，X 20，X 21，X 27，X 31和X 36。
TYYRDSVKGRFTISR X 16NAK X 20X 21LYLQM X 27SLR X 31EDTA X 36YYCST(SEQ ID NO：64)，其中，X 16可以是D或E，X 20可以是N或S，X 21可以是S或T，X 27可以是D或N，X 31可以是A或S，X 36可以是T或V。
在本申请中，所述抗原结合蛋白的H-FR3可包含SEQ ID NO:18和SEQ ID NO:29中任 一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR4可包含SEQ ID NO：65所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR4与SEQ ID NO：65所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 5和X 6。
WGQG X 5X 6VTVSS(SEQ ID NO：65)，其中，X 5可以是T或V，X 6可以是L或M。
在本申请中，所述抗原结合蛋白的H-FR4可包含SEQ ID NO:20和SEQ ID NO:30中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：62所示的氨基酸序列；所述H-FR2可包含SEQ ID NO：63所示的氨基酸序列；所述H-FR3可包含SEQ ID NO：64所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO：65所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO:14和SEQ ID NO:27中任一项所示的氨基酸序列；所述H-FR2可包含SEQ ID NO:16和SEQ ID NO:28中任一项所示的氨基酸序列；所述H-FR3可包含SEQ ID NO:18和SEQ ID NO:29中任一项所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO:20和SEQ ID NO:30中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：14所示的氨基酸序列；所述H-FR2可包含SEQ ID NO：16所示的氨基酸序列；所述H-FR3可包含SEQ ID NO：18所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO：20所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6或与其具有相同H-FR1-4的抗原结合片段。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：27所示的氨基酸序列；所述H-FR2可包含SEQ ID NO：28所示的氨基酸序列；所述H-FR3可包含SEQ ID NO：29所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO：30所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A63或与其具有相同H-FR1-4的抗原结合片段。
在本申请中，所述抗原结合蛋白可包含重链可变区，所述重链可变区可包含SEQ ID NO：67所示的氨基酸序列。例如，所述抗原结合蛋白包含VH，所述VH与SEQ ID NO：67所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 16，X 19，X 40，X 42，X 73，X 77，X 78，X 84，X 88，X 93，X 119，X 120。
EVQLVESGGGLVQPGX 16SLX 19LSCAASGFTFNDYNMAWVRQX 40PX 42KGLEWVATILFDGSRTYYRDSVKGRFTISRX 73NAKX 77X 78LYLQMX 84SLRX 88EDTAX 93YYCSTESPYYSEGYYQGYFDYWGQG X 119X 120VTVSS(SEQ ID NO：67)，其中，X 16可以是G或R，X 19可以是 R或S，X 40可以是A或T，X 42可以是G或K，X 73可以是D或E，X 77可以是N或S，X 78可以是S或T，X 84可以是D或N，X 88可以是A或S，X 93可以是T或V，X 119可以是T或V，X 120可以是L或M。
在本申请中，所述抗原结合蛋白的重链可变区可包含SEQ ID NO:13和SEQ ID NO:26中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包含重链恒定区，所述重链恒定区可包括源自IgG的恒定区或源自IgY的恒定区。
例如，所述抗原结合蛋白的重链恒定区可包含SEQ ID NO：56所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可以包含抗体轻链可变区VL中的至少一个CDR，所述VL可以包含SEQ ID NO：66或SEQ ID NO：76所示的氨基酸序列。
例如，所述VL可以包含SEQ ID NO：66或SEQ ID NO：76中任一项所示的氨基酸序列。在本申请中，所述分离的抗原结合蛋白的LCDR可以通过任何形式划分，只要VL与SEQ ID NO：66或SEQ ID NO：76中任一项所示的氨基酸序列相同，以任何形式划分得到的LCDR都可落入本申请的保护范围内。
在本申请中，所述抗原结合蛋白可包括轻链可变区VL，所述VL可包含LCDR1、LCDR2和LCDR3中的至少一个，至少两个或至少三个。
在本申请中，所述抗原结合蛋白的LCDR3可包含SEQ ID NO：11所示的氨基酸序列。例如，所述抗原结合蛋白的LCDR3可根据Chothia编号系统定义。
在本申请中，所述抗原结合蛋白的LCDR2可包含SEQ ID NO：9所示的氨基酸序列。例如，所述抗原结合蛋白的LCDR2可根据Chothia编号系统定义。
在本申请中，所述抗原结合蛋白的LCDR1可包含SEQ ID NO：7所示的氨基酸序列。例如，所述抗原结合蛋白的LCDR1可根据Chothia编号系统定义。
例如，本申请所述抗原结合蛋白的LCDR1可包含SEQ ID NO：7所示的氨基酸序列；所述LCDR2可包含SEQ ID NO：9所示的氨基酸序列；且所述LCDR3可包含SEQ ID NO：11所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6、JYB1931A63或与其具有相同LCDR3(例如，与其具有相同LCDR1-3)的抗原结合片段。
例如，所述抗原结合蛋白的VL可包含框架区L-FR1，L-FR2，L-FR3和L-FR4。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：58所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR1与SEQ ID NO：58所示的序列相比，在选自下组的一个或多 个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 1，X 13，X 14，X 18，X 19和X 20。
X 1IVLTQSPATLS X 13X 14PGE X 18X 19X 20LSC(SEQ ID NO：58)，其中，X 1可以是E或N，X 13可以是L或V，X 14可以是S或T，X 18可以是R或S，X 19可以是A或V，X 20可以是S或T。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO:6和SEQ ID NO:22中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR2可包含SEQ ID NO：59所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR2与SEQ ID NO：59所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 5，X 6，X 7，X 8和X 9。
WYQQ X 5X 6X 7X 8X 9PRLLIK(SEQ ID NO：59)，其中，X 5可以是K或R，X 6可以是P或S，X 7可以是G或N，X 8可以是E或Q，X 9可以是A或S。
在本申请中，所述抗原结合蛋白的L-FR2可包含SEQ ID NO:8和SEQ ID NO:23中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR3可包含SEQ ID NO：60所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR3与SEQ ID NO：60所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 4，X 18，X 20，X 21，X 22，X 24，X 28和X 29。
GIPX 4RFSGSGSGTDFTLX 18I X 20X 21X 22EX 24EDFX 28X 29YYC(SEQ ID NO：60)，其中，X 4可以是A或S，X 18可以是S或T，X 20可以是N或S，X 21可以是R或S，X 22可以是L或V，X 24可以是P或S，X 28可以是A或S，X 29可以是I或V。
在本申请中，所述抗原结合蛋白的L-FR3可包含SEQ ID NO:10和SEQ ID NO:24中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR4可包含SEQ ID NO：61所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR4与SEQ ID NO：61所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 3，X 9和X 10。
FG X 3GTKLE X 9X 10(SEQ ID NO：61)，其中，X 3可以是A或Q，X 9可以是I或L，X 10可以是K或R。
在本申请中，所述抗原结合蛋白的L-FR4可包含SEQ ID NO:12和SEQ ID NO:25中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：58所示的氨基酸序列；所述L-FR2可包含SEQ ID NO：59所示的氨基酸序列；所述L-FR3可包含SEQ ID NO：60所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO：61所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO:6和SEQ ID NO:22中任一项所示的氨基酸序列；所述L-FR2可包含SEQ ID NO:8和SEQ ID NO:23中任一项所示的氨基酸序列；所述L-FR3可包含SEQ ID NO:10和SEQ ID NO:24中任一项所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO:12和SEQ ID NO:25中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：6所示的氨基酸序列；所述L-FR2可包含SEQ ID NO：8所示的氨基酸序列；所述L-FR3可包含SEQ ID NO：10所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO：12所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6或与其具有相同L-FR1-4的抗体。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：22所示的氨基酸序列；所述L-FR2可包含SEQ ID NO：23所示的氨基酸序列；所述L-FR3可包含SEQ ID NO：24所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO：25所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A63或与其具有相同H-FR1-4的抗体。
在本申请中，所述抗原结合蛋白可包含轻链可变区VL，所述VL可包含SEQ ID NO：66所示的氨基酸序列。例如，所述抗原结合蛋白的VL与SEQ ID NO：66所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 1，X 13，X 14，X 18，X 19，X 20，X 39，X 40，X 41，X 42，X 43，X 60，X 74，X 76，X 77，X 78，X 80，X 84，X 85，X 100，X 106，X 107。
X 1IVLTQSPATLSX 13X 14PGEX 18X 19X 20LSCRASQSVSTSIHWYQQX 39X 40X 41X 42X 43PRLLIKYASRSISGIPX 60RFSGSGSGTDFTLX 74IX 76X 77X 78EX 80EDFX 84X 85YYCQQSYSSLYTFGX 100GTKLE X 106X 107(SEQ ID NO：66)，其中，X 1可以是E或N，X 13可以是L或V，X 14可以是S或T，X 18可以是R或S，X 19可以是A或V，X 20可以是S或T，X 39可以是K或R，X 40可以是P或S，X 41可以是G或N，X 42可以是E或Q，X 43可以是A或S，X 60可以是A或S，X 74可以是S或T，X 76可以是N或S，X 77可以是R或S，X 78可以是L或V，X 80可以是P或S，X 84可以是A或S，X 85可以是I或V，X 100可以是A或Q，X 106可以是I或L，X 107可以是K或R。
在本申请中，所述抗原结合蛋白的轻链可变区可包含SEQ ID NO:5和SEQ ID NO:21中 任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包含轻链恒定区，所述轻链恒定区可包括源自Igκ的恒定区或源自Igλ的恒定区。
例如，所述轻链恒定区可包括源自Igκ的恒定区。
例如，所述抗原结合蛋白的轻链恒定区包含SEQ ID NO：57所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包含HCDR1-3和LCDR1-3。例如，所述抗原结合蛋白的HCDR1可包含SEQ ID NO:15所示的氨基酸序列；所述抗原结合蛋白的HCDR2可包含SEQ ID NO:17所示的氨基酸序列；所述抗原结合蛋白的HCDR3可包含SEQ ID NO:19所示的氨基酸序列；所述抗原结合蛋白的LCDR1可包括SEQ ID NO:7所示的氨基酸序列；所述抗原结合蛋白的LCDR2可包含SEQ ID NO:9所示的氨基酸序列；所述抗原结合蛋白的LCDR3可包含SEQ ID NO:11所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6、JYB1931A63或与其具有相同HCDR3(例如，与其具有相同HCDR1-3)和LCDR3(例如，与其具有相同LCDR1-3)的抗原结合片段。
在本申请中，所述抗原结合蛋白可包含重链可变区和轻链可变区。所述抗原结合蛋白的重链可变区可包含HCDR1-3以及H-FR1-4。所述抗原结合蛋白的轻链可变区可包含LCDR1-3以及L-FR1-4。例如，所述HCDR1可包含SEQ ID NO:15所示的氨基酸序列；所述HCDR2可包含SEQ ID NO:17所示的氨基酸序列；所述HCDR3可包含SEQ ID NO:19所示的氨基酸序列；所述LCDR1可包括SEQ ID NO:7所示的氨基酸序列；所述LCDR2可包含SEQ ID NO:9所示的氨基酸序列；所述LCDR3可包含SEQ ID NO:11所示的氨基酸序列。例如，所述H-FR1可包含SEQ ID NO:14所示的氨基酸序列；所述H-FR2可包含SEQ ID NO:16所示的氨基酸序列；所述H-FR3可包含SEQ ID NO:18所示的氨基酸序列；所述H-FR4可包含SEQ ID NO:20所示的氨基酸序列；所述L-FR1可包括SEQ ID NO:6的氨基酸序列；所述L-FR2可包含SEQ ID NO:8所示的氨基酸序列；所述L-FR3可包含SEQ ID NO:10所示的氨基酸序列；所述L-FR4可包含SEQ ID NO:12所示的氨基酸序列。例如，所述抗原结合蛋白的重链可变区可包含SEQ ID NO:13所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6或与其具有相同重链可变区的抗原结合蛋白。例如，所述抗原结合蛋白的轻链可变区可包含SEQ ID NO:5所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体50A6或与其具有相同轻链可变区的抗原结合蛋白。
在本申请中，所述抗原结合蛋白可包含重链可变区和轻链可变区，所述重链可变区可包 含HCDR1-3以及H-FR1-4。所述轻链可变区可包含LCDR1-3以及L-FR1-4。例如，所述HCDR1可包含SEQ ID NO:15所示的氨基酸序列；所述HCDR2可包含SEQ ID NO:17所示的氨基酸序列；所述HCDR3可包含SEQ ID NO:19所示的氨基酸序列；所述LCDR1可包括SEQ ID NO:7所示的氨基酸序列；所述LCDR2可包含SEQ ID NO:9所示的氨基酸序列；所述LCDR3可包含SEQ ID NO:11所示的氨基酸序列。例如，所述H-FR1可包含SEQ ID NO:27所示的氨基酸序列；所述H-FR2可包含SEQ ID NO:28所示的氨基酸序列；所述H-FR3可包含SEQ ID NO:29所示的氨基酸序列；所述H-FR4可包含SEQ ID NO:30所示的氨基酸序列；所述L-FR1可包括SEQ ID NO:22的氨基酸序列；所述L-FR2可包含SEQ ID NO:23所示的氨基酸序列；所述L-FR3可包含SEQ ID NO:24所示的氨基酸序列；所述L-FR4可包含SEQ ID NO:25所示的氨基酸序列。例如，所述抗原结合蛋白的重链可变区可包含SEQ ID NO:26所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A63或与其具有相同重链可变区的抗原结合蛋白。例如，所述抗原结合蛋白的轻链可变区可包含SEQ ID NO:21所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A63或与其具有相同轻链可变区的抗原结合蛋白。
在本申请中，所述抗原结合蛋白可包含重链可变区VH，所述VH可包含HCDR1、HCDR2和HCDR3中的至少一个、两个或三个。
在本申请中，所述抗原结合蛋白的HCDR3可包含SEQ ID NO：45所示的氨基酸序列。例如，所述抗原结合蛋白的HCDR3序列可根据Chothia编码系统定义。
在本申请中，所述抗原结合蛋白的HCDR2可包含SEQ ID NO：43所示的氨基酸序列。例如，所述抗原结合蛋白的HCDR2序列可根据Chothia编码系统定义。
在本申请中，所述抗原结合蛋白的HCDR1可包含SEQ ID NO：41所示的氨基酸序列。例如，所述抗原结合蛋白的HCDR1序列可根据Chothia编码系统定义。
例如，所述抗原结合蛋白的HCDR1可包含SEQ ID NO：41所示的氨基酸序列；所述HCDR2可包含SEQ ID NO：43所示的氨基酸序列；且所述HCDR3可包含SEQ ID NO：45所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1、JYB1931A13或与其具有相同HCDR3(例如，与其具有相同的HCDR1-3)的抗原结合片段。
例如，所述抗原结合蛋白的VH可包含框架区H-FR1，H-FR2，H-FR3和H-FR4。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：72所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR1与SEQ ID NO：72所示的序列相比，在选自下组的一个或多 个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 1，X 17和X 25。
X 1VQLQESGPGLVKPSQ X 17LSLTCTV X 25(SEQ ID NO：72)，其中，X 1可以是D或Q，X 17可以是S或T，X 25可以是S或T。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO:40和SEQ ID NO:53中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR2可包含SEQ ID NO：73所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR2与SEQ ID NO：73所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 8，X 11，X 12和X 16。
AWNWIRQ X 8PG X 11X 12LEW X 16GYI(SEQ ID NO：73)，其中，X 8可以是F或P，X 11可以是K或N，X 12可以是G或K，X 16可以是I或M。
在本申请中，所述抗原结合蛋白的H-FR2可包含SEQ ID NO:42和SEQ ID NO:54中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR3可包含SEQ ID NO：74所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR3与SEQ ID NO：74所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 11，X 12，X 14，X 16，X 19，X 23，X 25，X 27，X 31，X 32，X 36和X 38。
TSYNPSLKSR X 11X 12I X 14R X 16TS X 19NQF X 23L X 25L X 27SVT X 31X 32DTA X 36Y X 38CAR(SEQ ID NO：74)，其中，X 11可以是I或V，X 12可以是S或T，X 14可以是S或T，X 16可以是D或N，X 19可以是K或T，X 23可以是F或S，X 25可以是K或Q，X 27可以是N或S，X 31可以是A或T，X 32可以是A或E，X 36可以是T或V，X 38可以是F或Y。
在本申请中，所述抗原结合蛋白的H-FR3可包含SEQ ID NO:44和SEQ ID NO:55中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR4可包含SEQ ID NO：75所示的氨基酸序列。例如，所述抗原结合蛋白的H-FR4与SEQ ID NO：75所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 6和X 7。
WGQGT X 6X 7TVSS(SEQ ID NO：75)，其中，X 6可以是L或T，X 7可以是L或V。
在本申请中，所述抗原结合蛋白的H-FR4可包含SEQ ID NO:46和SEQ ID NO:30中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：72所示的氨基酸序列；所 述H-FR2可包含SEQ ID NO：73所示的氨基酸序列；所述H-FR3可包含SEQ ID NO：74所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO：75所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO:40和SEQ ID NO:53中任一项所示的氨基酸序列；所述H-FR2可包含SEQ ID NO:42和SEQ ID NO:54中任一项所示的氨基酸序列；所述H-FR3可包含SEQ ID NO:44和SEQ ID NO:55中任一项所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO:46和SEQ ID NO:30中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：40所示的氨基酸序列；所述H-FR2可包含SEQ ID NO：42所示的氨基酸序列；所述H-FR3可包含SEQ ID NO：44所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO：46所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1或与其具有相同H-FR1-4的抗原结合片段。
在本申请中，所述抗原结合蛋白的H-FR1可包含SEQ ID NO：53所示的氨基酸序列；所述H-FR2可包含SEQ ID NO：54所示的氨基酸序列；所述H-FR3可包含SEQ ID NO：55所示的氨基酸序列；且所述H-FR4可包含SEQ ID NO：30所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A13或与其具有相同H-FR1-4的抗原结合片段。
在本申请中，所述抗原结合蛋白可包含重链可变区，所述重链可变区可包含SEQ ID NO：77所示的氨基酸序列。例如，所述抗原结合蛋白包含VH，所述VH与SEQ ID NO：77所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 1，X 17，X 25，X 41，X 44，X 45，X 49，X 68，X 69，X 71，X 73，X 76，X 80，X 82，X 84，X 88，X 89，X 93，X 95，X 109和X 110。
X 1VQLQESGPGLVKPSQ X 17LSLTCTV X 25GYSITSDYAWNWIRQ X 41PG X 44X 45LEWX 49GYISYSGRTSYNPSLKSR X 68X 69I X 71R X 73TS X 76NQF X 80L X 82L X 84SVT X 88X 89DTA X 93YX 95CARYWGDYWGQGT X 109X 110TVSS(SEQ ID NO：77)，其中，X 1可以是D或Q，X 17可以是S或T，X 25可以是S或T，X 41可以是F或P，X 44可以是K或N，X 45可以是G或K，X 49可以是I或M，X 68可以是I或V，X 69可以是S或T，X 71可以是S或T，X 73可以是D或N，X 76可以是K或T，X 80可以是F或S，X 82可以是K或Q，X 84可以是N或S，X 88可以是A或T，X 89可以是A或E，X 93可以是T或V，X 95可以是F或Y，X 109可以是L或T，X 110可以是L或V。
在本申请中，所述抗原结合蛋白的重链可变区可包含SEQ ID NO:39和SEQ ID NO:52中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包含重链恒定区，所述重链恒定区可包括源自IgG的恒定区或源自IgY的恒定区。
例如，所述抗原结合蛋白的重链恒定区可包含SEQ ID NO：56所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包括轻链可变区VL，所述VL可包含LCDR1、LCDR2和LCDR3中的至少一个，至少两个或至少三个。
在本申请中，所述抗原结合蛋白的LCDR3可包含SEQ ID NO：37所示的氨基酸序列。例如，所述抗原结合蛋白的LCDR3可根据Chothia编号系统定义。
在本申请中，所述抗原结合蛋白的LCDR2可包含SEQ ID NO：35所示的氨基酸序列。例如，所述抗原结合蛋白的LCDR2可根据Chothia编号系统定义。
在本申请中，所述抗原结合蛋白的LCDR1可包含SEQ ID NO：33所示的氨基酸序列。例如，所述抗原结合蛋白的LCDR1可根据Chothia编号系统定义。
例如，本申请所述抗原结合蛋白的LCDR1可包含SEQ ID NO：33所示的氨基酸序列；所述LCDR2可包含SEQ ID NO：35所示的氨基酸序列；且所述LCDR3可包含SEQ ID NO：37所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1、JYB1931A13或与其具有相同LCDR3(例如，与其具有相同LCDR1-3)的抗原结合片段。
例如，所述抗原结合蛋白的VL可包含框架区L-FR1，L-FR2，L-FR3和L-FR4。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：68所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR1与SEQ ID NO：68所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 3，X 8，X 9，X 10，X 11，X 13，X 20和X 21。
DI X 3MTQS X 8X 9X 10X 11S X 13SVGDRV X 20X 21TC(SEQ ID NO：68)，其中，X 3可以是Q或V，X 8可以是P或Q，X 9可以是K或S，X 10可以是F或S，X 11可以是L或M，X 13可以是A或T，X 20可以是S或T，X 21可以是I或V。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO:32和SEQ ID NO:48中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR2可包含SEQ ID NO：69所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR2与SEQ ID NO：69所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 8和X 9。
WFQQKPG X 8X 9PKPLIY(SEQ ID NO：69)，其中，X 8可以是K或Q，X 9可以是A或 S。
在本申请中，所述抗原结合蛋白的L-FR2可包含SEQ ID NO:34和SEQ ID NO:49中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR3可包含SEQ ID NO：70所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR3与SEQ ID NO：70所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 4，X 7，X 21，X 22，X 24，X 27和X 29。
GVP X 4RF X 7GSGSGTDFTLTIS X 21X 22Q X 24ED X 27A X 29YFC(SEQ ID NO：70)，其中，X 4可以是D或S，X 7可以是S或T，X 21可以是N或S，X 22可以是L或V，X 24可以是P或S，X 27可以是F或L，X 29可以是E或T。
在本申请中，所述抗原结合蛋白的L-FR3可包含SEQ ID NO:36和SEQ ID NO:50中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR4可包含SEQ ID NO：71所示的氨基酸序列。例如，所述抗原结合蛋白的L-FR4与SEQ ID NO：71所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 3，X 7，X 9，X 10。
FG X 3GTK X 7E X 9X 10(SEQ ID NO：71)，其中，X 3可以是A或G，X 7可以是L或V，X 9可以是I或L，X 10可以是K或N。
在本申请中，所述抗原结合蛋白的L-FR4可包含SEQ ID NO:38和SEQ ID NO:51中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：68所示的氨基酸序列；所述L-FR2可包含SEQ ID NO：69所示的氨基酸序列；所述L-FR3可包含SEQ ID NO：70所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO：71所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO:32和SEQ ID NO:48中任一项所示的氨基酸序列；所述L-FR2可包含SEQ ID NO:34和SEQ ID NO:49中任一项所示的氨基酸序列；所述L-FR3可包含SEQ ID NO:36和SEQ ID NO:50中任一项所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO:38和SEQ ID NO:51中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：32所示的氨基酸序列；所述L-FR2可包含SEQ ID NO：34所示的氨基酸序列；所述L-FR3可包含SEQ ID NO：36所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO：38所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1或与其具有相同L-FR1-4的抗体。
在本申请中，所述抗原结合蛋白的L-FR1可包含SEQ ID NO：48所示的氨基酸序列；所述L-FR2可包含SEQ ID NO：49所示的氨基酸序列；所述L-FR3可包含SEQ ID NO：50所示的氨基酸序列；且所述L-FR4可包含SEQ ID NO：51所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A13或与其具有相同H-FR1-4的抗体。
在本申请中，所述抗原结合蛋白可包含轻链可变区VL，所述VL可包含SEQ ID NO：76所示的氨基酸序列。例如，所述抗原结合蛋白的VL与SEQ ID NO：76所示的序列相比，在选自下组的一个或多个氨基酸处存在氨基酸取代(例如，保守氨基酸取代等)：X 3，X 8，X 9，X 10，X 11，X 13，X 20，X 21，X 42，X 43，X 60，X 63，X 77，X 78，X 80，X 83，X 85，X 100，X 104，X 106，X 107。
DIX 3MTQSX 8X 9X 10X 11SX 13SVGDRVX 20X 21TCKASQNVGSNVAWFQQKPGX 42X 43PKPLIYSASYRYSGVPX 60RFX 63GSGSGTDFTLTISX 77X 78QX 80EDX 83AX 85YFCHQYNTYPLTFGX 100GTK X 104EX 106X 107(SEQ ID NO：76)，其中，X 3可以是Q或V，X 8可以是P或Q，X 9可以是K或S，X 10可以是F或S，X 11可以是L或M，X 13可以是A或T，X 20可以是S或T，X 21可以是I或V，X 42可以是K或Q，X 43可以是A或S，X 60可以是D或S，X 63可以是S或T，X 77可以是N或S，X 78可以是L或V，X 80可以是P或S，X 83可以是F或L，X 85可以是E或T，X 100可以是A或G，X 104可以是L或V，X 106可以是I或L，X 107可以是K或N。
在本申请中，所述抗原结合蛋白的轻链可变区可包含SEQ ID NO：31和47中任一项所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包含轻链恒定区，所述轻链恒定区可包括源自Igκ的恒定区或源自Igλ的恒定区。
例如，所述轻链恒定区可包括源自Igκ的恒定区。
例如，所述抗原结合蛋白的轻链恒定区包含SEQ ID NO：57所示的氨基酸序列。
在本申请中，所述抗原结合蛋白可包含HCDR1-3和LCDR1-3。例如，所述抗原结合蛋白的HCDR1可包含SEQ ID NO:41所示的氨基酸序列；所述抗原结合蛋白的HCDR2可包含SEQ ID NO:43所示的氨基酸序列；所述抗原结合蛋白的HCDR3可包含SEQ ID NO:45所示的氨基酸序列；所述抗原结合蛋白的LCDR1可包括SEQ ID NO:33所示的氨基酸序列；所述抗原结合蛋白的LCDR2可包含SEQ ID NO:35所示的氨基酸序列；所述抗原结合蛋白的LCDR3可包含SEQ ID NO:37所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1、JYB1931A13或与其具有相同HCDR3(例如，与其具有相同HCDR1-3)和LCDR3(例如， 与其具有相同LCDR1-3)的抗原结合片段。
在本申请中，所述抗原结合蛋白可包含重链可变区和轻链可变区。所述抗原结合蛋白的重链可变区可包含HCDR1-3以及H-FR1-4。所述抗原结合蛋白的轻链可变区可包含LCDR1-3以及L-FR1-4。例如，所述HCDR1可包含SEQ ID NO:41所示的氨基酸序列；所述HCDR2可包含SEQ ID NO:43所示的氨基酸序列；所述HCDR3可包含SEQ ID NO:45所示的氨基酸序列；所述LCDR1可包括SEQ ID NO:33所示的氨基酸序列；所述LCDR2可包含SEQ ID NO:35所示的氨基酸序列；所述LCDR3可包含SEQ ID NO:37所示的氨基酸序列。例如，所述H-FR1可包含SEQ ID NO:40所示的氨基酸序列；所述H-FR2可包含SEQ ID NO:42所示的氨基酸序列；所述H-FR3可包含SEQ ID NO:44所示的氨基酸序列；所述H-FR4可包含SEQ ID NO:46所示的氨基酸序列；所述L-FR1可包括SEQ ID NO:32的氨基酸序列；所述L-FR2可包含SEQ ID NO:34所示的氨基酸序列；所述L-FR3可包含SEQ ID NO:36所示的氨基酸序列；所述L-FR4可包含SEQ ID NO:38所示的氨基酸序列。例如，所述抗原结合蛋白的重链可变区可包含SEQ ID NO:39所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1或与其具有相同重链可变区的抗原结合蛋白。例如，所述抗原结合蛋白的轻链可变区可包含SEQ ID NO:31所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体47A1或与其具有相同轻链可变区的抗原结合蛋白。
在本申请中，所述抗原结合蛋白可包含重链可变区和轻链可变区，所述重链可变区可包含HCDR1-3以及H-FR1-4。所述轻链可变区可包含LCDR1-3以及L-FR1-4。例如，所述HCDR1可包含SEQ ID NO:41所示的氨基酸序列；所述HCDR2可包含SEQ ID NO:43所示的氨基酸序列；所述HCDR3可包含SEQ ID NO:45所示的氨基酸序列；所述LCDR1可包括SEQ ID NO:33所示的氨基酸序列；所述LCDR2可包含SEQ ID NO:35所示的氨基酸序列；所述LCDR3可包含SEQ ID NO:37所示的氨基酸序列。例如，所述H-FR1可包含SEQ ID NO:53所示的氨基酸序列；所述H-FR2可包含SEQ ID NO:54所示的氨基酸序列；所述H-FR3可包含SEQ ID NO:55所示的氨基酸序列；所述H-FR4可包含SEQ ID NO:30所示的氨基酸序列；所述L-FR1可包括SEQ ID NO:48的氨基酸序列；所述L-FR2可包含SEQ ID NO:49所示的氨基酸序列；所述L-FR3可包含SEQ ID NO:50所示的氨基酸序列；所述L-FR4可包含SEQ ID NO:51所示的氨基酸序列。例如，所述抗原结合蛋白的重链可变区可包含SEQ ID NO:52所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A13或与其具有相同重链可变区的抗原结合蛋白。例如，所述抗原结合蛋白的轻链可变区可包含SEQ ID NO: 47所示的氨基酸序列。例如，所述抗原结合蛋白可包括抗体JYB1931A13或与其具有相同轻链可变区的抗原结合蛋白。
在本申请中，所述分离的抗原结合蛋白还可与参比抗体竞争结合所述人MASP-2蛋白，所述参比抗体可包含重链可变区VH，所述VH可包含HCDR1、HCDR2和HCDR3中的至少一个、两个或三个。
在本申请中，所述参比抗体的HCDR3可包含SEQ ID NO：19所示的氨基酸序列。例如，所述参比抗体的HCDR3的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的HCDR2可包含SEQ ID NO：17所示的氨基酸序列。例如，所述参比抗体的HCDR2的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的HCDR1可包含SEQ ID NO：15所示的氨基酸序列。例如，所述参比抗体的HCDR1的序列可根据Chothia编码系统定义。
例如，所述参比抗体的HCDR1可包含SEQ ID NO：15所示的氨基酸序列；所述HCDR2可包含SEQ ID NO：17所示的氨基酸序列；且所述HCDR3可包含SEQ ID NO：19所示的氨基酸序列。例如，所述参比抗体可包括抗体50A6、JYB1931A63或与其具有相同HCDR3(例如，与其具有相同HCDR1-3)的抗原结合蛋白。
在本申请中，所述参比抗体可包含重链可变区，所述重链可变区可包含SEQ ID NO：67所示的氨基酸序列。
EVQLVESGGGLVQPGX 16SLX 19LSCAASGFTFNDYNMAWVRQX 40PX 42KGLEWVATILFDGSRTYYRDSVKGRFTISRX 73NAKX 77X 78LYLQMX 84SLRX 88EDTAX 93YYCSTESPYYSEGYYQGYFDYWGQG X 119X 120VTVSS(SEQ ID NO：67)，其中，X 16可以是G或R，X 19可以是R或S，X 40可以是A或T，X 42可以是G或K，X 73可以是D或E，X 77可以是N或S，X 78可以是S或T，X 84可以是D或N，X 88可以是A或S，X 93可以是T或V，X 119可以是T或V，X 120可以是L或M。
在本申请中，所述参比抗体的重链可变区可包含SEQ ID NO:13和SEQ ID NO:26中任一项所示的氨基酸序列。
在本申请中，所述参比抗体可包含重链恒定区，所述重链恒定区可包括源自IgG的恒定区或源自IgY的恒定区。
例如，所述参比抗体的重链恒定区可包含SEQ ID NO：56所示的氨基酸序列。
在本申请中，所述参比抗体可包括轻链可变区VL，所述VL可包含LCDR1、LCDR2和 LCDR3。
在本申请中，所述参比抗体的LCDR3可包含SEQ ID NO：11所示的氨基酸序列。例如，所述参比抗体的LCDR3的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的LCDR2可包含SEQ ID NO：9所示的氨基酸序列。例如，所述参比抗体的LCDR2的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的LCDR1可包含SEQ ID NO：7所示的氨基酸序列。例如，所述参比抗体的LCDR1的序列可根据Chothia编码系统定义。
例如，本申请所述参比抗体的LCDR1可包含SEQ ID NO：7所示的氨基酸序列；所述LCDR2可包含SEQ ID NO：9所示的氨基酸序列；且所述LCDR3可包含SEQ ID NO：11所示的氨基酸序列。例如，所述参比抗体可包括抗体50A6、JYB1931A63或与其具有相同LCDR3(例如，与其具有相同LCDR1-3)的抗原结合蛋白。
在本申请中，所述参比抗体可包含轻链可变区，所述轻链可变区可包含SEQ ID NO：66所示的氨基酸序列。
X 1IVLTQSPATLSX 13X 14PGEX 18X 19X 20LSCRASQSVSTSIHWYQQX 39X 40X 41X 42X 43PRLLIKYASRSISGIPX 60RFSGSGSGTDFTLX 74IX 76X 77X 78EX 80EDFX 84X 85YYCQQSYSSLYTFGX 100GTKLE X 106X 107(SEQ ID NO：66)，其中，X 1可以是E或N，X 13可以是L或V，X 14可以是S或T，X 18可以是R或S，X 19可以是A或V，X 20可以是S或T，X 39可以是K或R，X 40可以是P或S，X 41可以是G或N，X 42可以是E或Q，X 43可以是A或S，X 60可以是A或S，X 74可以是S或T，X 76可以是N或S，X 77可以是R或S，X 78可以是L或V，X 80可以是P或S，X 84可以是A或S，X 85可以是I或V，X 100可以是A或Q，X 106可以是I或L，X 107可以是K或R。
在本申请中，所述参比抗体的轻链可变区可包含SEQ ID NO:5和SEQ ID NO:21中任一项所示的氨基酸序列。
在本申请中，所述参比抗体可包含HCDR1-3和LCDR1-3。例如，所述HCDR1可包含SEQ ID NO:15所示的氨基酸序列；所述HCDR2可包含SEQ ID NO:17所示的氨基酸序列；所述HCDR3可包含SEQ ID NO:19所示的氨基酸序列；所述LCDR1可包括SEQ ID NO:7所示的氨基酸序列；所述LCDR2可包含SEQ ID NO:9所示的氨基酸序列；所述LCDR3可包含SEQ ID NO:11所示的氨基酸序列。例如，所述参比抗体可包括抗体50A6、JYB1931A63或与其具有相同HCDR3(例如，与其具有相同HCDR1-3)和LCDR3(例如，与其具有相同 LCDR1-3)的抗原结合蛋白。
在本申请中，所述分离的抗原结合蛋白还可与参比抗体竞争结合所述人MASP-2蛋白，所述参比抗体可包含重链可变区VH，所述VH可包含HCDR1、HCDR2和HCDR3中的至少一个、两个或三个。
在本申请中，所述参比抗体的HCDR3可包含SEQ ID NO：45所示的氨基酸序列。例如，所述参比抗体的HCDR3的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的HCDR2可包含SEQ ID NO：43所示的氨基酸序列。例如，所述参比抗体的HCDR2的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的HCDR1可包含SEQ ID NO：41所示的氨基酸序列。例如，所述参比抗体的HCDR1的序列可根据Chothia编码系统定义。
例如，所述参比抗体的HCDR1可包含SEQ ID NO：41所示的氨基酸序列；所述HCDR2可包含SEQ ID NO：43所示的氨基酸序列；且所述HCDR3可包含SEQ ID NO：45所示的氨基酸序列。例如，所述参比抗体可包括抗体47A1、JYB1931A13或与其具有相同HCDR3(例如，与其具有相同HCDR1-3)的抗原结合蛋白。
在本申请中，所述参比抗体可包含重链可变区，所述重链可变区可包含SEQ ID NO：77所示的氨基酸序列。
X 1VQLQESGPGLVKPSQ X 17LSLTCTV X 25GYSITSDYAWNWIRQ X 41PG X 44X 45LEWX 49GYISYSGRTSYNPSLKSR X 68X 69I X 71R X 73TS X 76NQF X 80L X 82L X 84SVT X 88X 89DTA X 93YX 95CARYWGDYWGQGT X 109X 110TVSS(SEQ ID NO：77)，其中，X 1可以是D或Q，X 17可以是S或T，X 25可以是S或T，X 41可以是F或P，X 44可以是K或N，X 45可以是G或K，X 49可以是I或M，X 68可以是I或V，X 69可以是S或T，X 71可以是S或T，X 73可以是D或N，X 76可以是K或T，X 80可以是F或S，X 82可以是K或Q，X 84可以是N或S，X 88可以是A或T，X 89可以是A或E，X 93可以是T或V，X 95可以是F或Y，X 109可以是L或T，X 110可以是L或V。
在本申请中，所述参比抗体的重链可变区可包含SEQ ID NO:39和SEQ ID NO:52中任一项所示的氨基酸序列。
在本申请中，所述参比抗体可包含重链恒定区，所述重链恒定区可包括源自IgG的恒定区或源自IgY的恒定区。
例如，所述参比抗体的重链恒定区可包含SEQ ID NO：56所示的氨基酸序列。
在本申请中，所述参比抗体可包括轻链可变区VL，所述VL可包含LCDR1、LCDR2和LCDR3。
在本申请中，所述参比抗体的LCDR3可包含SEQ ID NO：37所示的氨基酸序列。例如，所述参比抗体的LCDR3的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的LCDR2可包含SEQ ID NO：35所示的氨基酸序列。例如，所述参比抗体的LCDR2的序列可根据Chothia编码系统定义。
在本申请中，所述参比抗体的LCDR1可包含SEQ ID NO：33所示的氨基酸序列。例如，所述参比抗体的LCDR1的序列可根据Chothia编码系统定义。
例如，本申请所述参比抗体的LCDR1可包含SEQ ID NO：33所示的氨基酸序列；所述LCDR2可包含SEQ ID NO：35所示的氨基酸序列；且所述LCDR3可包含SEQ ID NO：37所示的氨基酸序列。例如，所述参比抗体可包括抗体47A1、JYB1931A13或与其具有相同LCDR3(例如，与其具有相同LCDR1-3)的抗原结合蛋白。
在本申请中，所述参比抗体可包含轻链可变区，所述轻链可变区可包含SEQ ID NO：76所示的氨基酸序列。
DIX 3MTQSX 8X 9X 10X 11SX 13SVGDRVX 20X 21TCKASQNVGSNVAWFQQKPGX 42X 43PKPLIYSASYRYSGVPX 60RFX 63GSGSGTDFTLTISX 77X 78QX 80EDX 83AX 85YFCHQYNTYPLTFGX 100GTK X 104EX 106X 107(SEQ ID NO：76)，其中，X 3可以是Q或V，X 8可以是P或Q，X 9可以是K或S，X 10可以是F或S，X 11可以是L或M，X 13可以是A或T，X 20可以是S或T，X 21可以是I或V，X 42可以是K或Q，X 43可以是A或S，X 60可以是D或S，X 63可以是S或T，X 77可以是N或S，X 78可以是L或V，X 80可以是P或S，X 83可以是F或L，X 85可以是E或T，X 100可以是A或G，X 104可以是L或V，X 106可以是I或L，X 107可以是K或N。
在本申请中，所述参比抗体的轻链可变区可包含SEQ ID NO：31和47中任一项所示的氨基酸序列。
在本申请中，所述参比抗体可包含HCDR1-3和LCDR1-3。例如，所述HCDR1可包含SEQ ID NO:41所示的氨基酸序列；所述HCDR2可包含SEQ ID NO:43所示的氨基酸序列；所述HCDR3可包含SEQ ID NO:45所示的氨基酸序列；所述LCDR1可包括SEQ ID NO:33所示的氨基酸序列；所述LCDR2可包含SEQ ID NO:35所示的氨基酸序列；所述LCDR3可包含SEQ ID NO:37所示的氨基酸序列。例如，所述参比抗体可包括抗体47A1、JYB1931A13或与其具有相同HCDR3(例如，与其具有相同HCDR1-3)和LCDR3(例如，与其具有相同 LCDR1-3)的抗原结合蛋白。
在本申请中，所述参比抗体可包含重链可变区和轻链可变区。例如，所述参比抗体的重链可变区可包含SEQ ID NO:13所示的氨基酸序列。例如，所述参比抗体可包括抗体50A6或与其具有相同重链可变区的抗原结合蛋白。例如，所述参比抗体的轻链可变区可包含SEQ ID NO:5所示的氨基酸序列。例如，所述参比抗体可包括抗体50A6或与其具有相同轻链可变区的抗原结合蛋白。例如，所述参比抗体可包括抗体50A6或与其具有相同重链可变区及轻链可变区的抗原结合蛋白。
在本申请中，所述参比抗体可包含重链可变区和轻链可变区。例如，所述参比抗体的重链可变区可包含SEQ ID NO:26所示的氨基酸序列。例如，所述参比抗体可包括抗体JYB1931A63或与其具有相同重链可变区的抗原结合蛋白。例如，所述参比抗体的轻链可变区可包含SEQ ID NO:21所示的氨基酸序列。例如，所述参比抗体可包括抗体JYB1931A63或与其具有相同轻链可变区的抗原结合蛋白。例如，所述参比抗体可包括抗体JYB1931A63或与其具有相同重链可变区及轻链可变区的抗原结合蛋白。
在本申请中，所述参比抗体可包含重链可变区和轻链可变区。例如，所述参比抗体的重链可变区可包含SEQ ID NO:39所示的氨基酸序列。例如，所述参比抗体可包括抗体47A1或与其具有相同重链可变区的抗原结合蛋白。例如，所述参比抗体的轻链可变区可包含SEQ ID NO:31所示的氨基酸序列。例如，所述参比抗体可包括抗体47A1或与其具有相同轻链可变区的抗原结合蛋白。例如，所述参比抗体可包括抗体47A1或与其具有相同重链可变区及轻链可变区的抗原结合蛋白。
在本申请中，所述参比抗体可包含重链可变区和轻链可变区。例如，所述参比抗体的重链可变区可包含SEQ ID NO:52所示的氨基酸序列。例如，所述参比抗体可包括抗体JYB1931A13或与其具有相同重链可变区的抗原结合蛋白。例如，所述参比抗体的轻链可变区可包含SEQ ID NO:47所示的氨基酸序列。例如，所述参比抗体可包括抗体JYB1931A13或与其具有相同轻链可变区的抗原结合蛋白。例如，所述参比抗体可包括抗体JYB1931A13或与其具有相同重链可变区及轻链可变区的抗原结合蛋白。
多肽和免疫缀合物
另一方面，本申请提供了一种或多种多肽，其可包含本申请的分离的抗原结合蛋白。例如，所述多肽可包括融合蛋白。例如，所述多肽可包括多特异性抗体(例如，双特异性抗体)。
另一方面，本申请提供了一种或多种免疫缀合物，所述免疫缀合物可包含本申请的分离 的抗原结合蛋白。在某些实施方式中，所述免疫缀合物还可包含药学上可接受的治疗剂、标记物和/或检测剂。
核酸、载体和细胞
另一方面，本申请还提供了分离的一种或多种核酸分子，所述一种或多种核酸分子可编码本申请所述分离的抗原结合蛋白。例如，所述一种或多种核酸分子中的每一个核酸分子可以编码完整的所述抗原结合蛋白，也可以编码其中的一部分(例如，HCDR1-3、重链可变区中的一种或多种)。
例如，当核酸分子分别编码所述抗原结合蛋白的一部分时，核酸分子编码的产物合在一起可以形成有功能的(例如，可结合MASP-2)的本申请的分离的抗原结合蛋白。
本申请所述的核酸分子可以为分离的。例如，其可以是通过以下方法产生或合成的：(i)在体外扩增的，例如通过聚合酶链式反应(PCR)扩增产生的，(ii)通过克隆重组产生的，(iii)纯化的，例如通过酶切和凝胶电泳分级分离，或者(iv)合成的，例如通过化学合成。例如，所述分离的核酸可以是通过重组DNA技术制备的核酸分子。
在本申请中，可以通过本领域已知的多种方法来制备编码所述分离的抗原结合蛋白的核酸，这些方法包括但不限于，采用逆转录PCR和PCR获得本申请所述分离的抗原结合蛋白的核酸分子。
另一方面，本申请提供了一种或多种载体，其包含本申请所述的一种或多种核酸分子。每种载体中可包含一种或多种所述核酸分子。此外，所述载体中还可包含其他基因，例如允许在适当的宿主细胞中和在适当的条件下选择该载体的标记基因。此外，所述载体还可包含允许编码区在适当宿主中正确表达的表达控制元件。这样的控制元件为本领域技术人员所熟知的，例如，可包括启动子、核糖体结合位点、增强子和调节基因转录或mRNA翻译的其他控制元件等。在某些实施方式中，所述表达控制序列为可调的元件。所述表达控制序列的具体结构可根据物种或细胞类型的功能而变化，但通常包含分别参与转录和翻译起始的5’非转录序列和5’及3’非翻译序列，例如TATA盒、加帽序列、CAAT序列等。例如，5’非转录表达控制序列可包含启动子区，启动子区可包含用于转录控制功能性连接核酸的启动子序列。所述表达控制序列还可包括增强子序列或上游活化子序列。在本申请中，适当的启动子可包括，例如用于SP6、T3和T7聚合酶的启动子、人U6RNA启动子、CMV启动子及其人工杂合启动子(如CMV)，其中启动子的某部分可与其他细胞蛋白(如人GAPDH，甘油醛-3-磷酸脱氢酶)基因启动子的某部分融合，其可包含或不包含另外的内含子。本申请所述的一种或多 种核酸分子可以与所述表达控制元件可操作地连接。
所述载体可以包括，例如质粒、粘粒、病毒、噬菌体或者在例如遗传工程中通常使用的其他载体。例如，所述载体可为表达载体。例如，所述载体可为病毒载体。可以将病毒载体直接给予至患者(体内)或可以通过间接的形式，例如，在体外使用病毒处理细胞，然后将处理过的细胞给予至患者(离体)。病毒载体技术在本领域中是公知的，并在例如Sambrook等(2001,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York)和其他病毒学和分子生物学手册中进行了描述。常规的基于病毒的系统可以包括用于基因转移的逆转录病毒载体、慢病毒载体、腺病毒载体、腺相关病毒载体以及单纯疱疹病毒载体。在某些情形中，可以用逆转录病毒、慢病毒和腺相关病毒的方法将基因转移整合进宿主基因组中，使插入的基因长期表达。慢病毒载体是能够转导或感染非分裂细胞并典型地产生较高病毒效价的逆转录病毒载体。慢病毒载体可包含长末端重复序列5’LTR和截短的3’LTR、RRE、rev应答元件(cPPT)、中央终止序列(CTS)和/或翻译后调控元件(WPRE)。本申请所述的载体可以被引入细胞。
另一方面，本申请提供了一种细胞。所述细胞可包含本申请所述的分离的抗原结合蛋白、所述的多肽、所述的免疫缀合物、一种或多种核酸分子和/或本申请所述的一种或多种载体。例如，每种或每个细胞可包含一个或一种本申请所述的核酸分子或载体。例如，每种或每个细胞可包含多个(例如，2个或以上)或多种(例如，2种或以上)本申请所述的核酸分子或载体。例如，可将本申请所述的载体引入所述宿主细胞中，例如原核细胞(例如，细菌细胞)、CHO细胞、NS/0细胞、HEK293T细胞、293F细胞或HEK293A细胞，或者其他真核细胞，如来自植物的细胞、真菌或酵母细胞等。可通过本领域已知的方法将本申请所述的载体引入所述宿主细胞中，例如电穿孔、lipofectine转染、lipofectamin转染等。例如，所述细胞可以包括酵母细胞。例如，所述细胞可以包括大肠杆菌细胞。例如，所述细胞可以包括哺乳动物细胞。例如，所述细胞可以包括免疫细胞。
所述细胞可以包括免疫细胞。在某些情形中，所述细胞可以包括免疫细胞。例如，所述细胞可包括T细胞、B细胞、天然杀伤(NK)细胞、巨噬细胞、NKT细胞、单核细胞、树突状细胞、粒细胞、淋巴细胞、白细胞和/或外周血单个核细胞。
药物组合物
另一方面，本申请提供了一种药物组合物。所述药物组合物可包含本申请所述的分离的抗原结合蛋白、所述的多肽、所述的免疫缀合物、所述分离的核酸分子、所述的载体、所述 的细胞，和/或药学上可接受的佐剂和/或赋形剂。在本申请中，所述药学上可接受的佐剂可以包括缓冲剂、抗氧化剂、防腐剂、低分子量多肽、蛋白质、亲水聚合物、氨基酸、糖、螯合剂、反离子、金属复合物和/或非离子表面活性剂。除非与本申请所述的细胞不相容，否则任何常规介质或试剂均可以考虑用于本申请的药物组合物中。在本申请中，所述药学上可接受的赋形剂可以包括在药物制剂中除主药以外的附加物，也可称为辅料。例如，所述赋形剂可以包括片剂中的粘合剂、填充剂、崩解剂、润滑剂。例如，所述赋形剂可以包括中药丸剂中的酒、醋、药汁等。例如，所述赋形剂可以包括半固体制剂软膏剂、霜剂中的基质部分。例如，所述赋形剂可以包括液体制剂中的防腐剂、抗氧剂、矫味剂、芳香剂、助溶剂、乳化剂、增溶剂、渗透压调节剂、着色剂。
试剂盒、用途和方法
另一方面，本申请提供了一种用于检测或测定MASP-2的方法，所述方法可包括使用所述分离的抗原结合蛋白或所述的多肽。
在本申请中，所述方法可包括体外方法，离体方法，非诊断或非治疗目的的方法。
例如，所述方法可包括用于非诊断目的的检测MASP-2的存在和/或含量的方法，其可包括下述步骤：
1)使样品与本申请的抗原结合蛋白接触；以及
2)检测样品结合的所述抗原结合蛋白的存在和/或含量来确定获自受试者的样品中MASP-2的存在和/或表达水平。
另一方面，本申请提供了一种MASP-2的试剂盒，其可包括使用所述分离的抗原结合蛋白或所述的多肽。
在本申请中，所述试剂盒还可包含使用说明，所述使用说明记载用于检测MASP-2的存在和/或含量的方法。例如，所述方法可包括体外方法，离体方法，非诊断或非治疗目的的方法。
另一方面，本申请提供了一种所述的分离的抗原结合蛋白或所述的多肽在制备试剂盒中的用途，所述试剂盒可用于检测MASP-2的存在和/或含量的方法。例如，所述方法可包括体外方法，离体方法，非诊断或非治疗目的的方法。
另一方面，本申请提供了分离的抗原结合蛋白、所述的多肽、所述的免疫缀合物、所述分离的核酸分子、所述的载体、所述的药物组合物用于预防、缓解和/或治疗疾病或病症。
另一方面，本申请提供了一种所述的分离的抗原结合蛋白、所述的多肽、所述的免疫缀 合物、所述的分离的核酸分子、所述的载体，所述的细胞和/或所述的药物组合物在制备药物中的用途，所述药物用于预防、缓解和/或治疗疾病或病症。
另一方面，本申请提供了一种预防和/或治疗疾病或病症的方法，其包括向有需要的受试者施用所述的分离的抗原结合蛋白、所述的分离的核酸分子、所述的载体，所述的细胞、所述的药物组合物。
本申请所述药物组合物、药物组合及方法可与其他类型的癌症疗法结合使用，诸如化学疗法、手术、放射、基因疗法等。本申请中所描述的药物组合物及方法可用于其他依赖于免疫反应的疾病病状，诸如炎症、免疫疾病及感染性疾病。
在本申请中，所述受试者可以包括人或非人动物。例如，所述非人动物可以选自下组：猴、鸡、鹅、猫、狗、小鼠和大鼠。此外，非人动物也可以包括任何除人以外的动物物种，例如家畜动物，或啮齿类动物，或灵长类动物，或家养动物，或家禽动物。所述人可以是高加索人、非洲人、亚洲人、闪族人，或其他种族，或各种种族的杂合体。又例如，所述人可以是老年、成年、青少年、儿童或者婴儿。
可以根据在实验动物中的有效量推测在人类中的有效量。例如，Freireich等人描述了动物和人的剂量的相互关系(基于每平方米身体表面的毫克数)(Freireich et al.，Cancer Chemother.Rep.50，219(1966))。身体表面积可以从患者的身高和体重近似确定。参见例如Scientific Tables，Geigy Pharmaceuticals，Ardsley，N.Y.，537(1970)。
不欲被任何理论所限，下文中的实施例仅仅是为了阐释本申请的融合蛋白、制备方法和用途等，而不用于限制本申请发明的范围。
实施例
实施例1抗原制备
人MASP-2蛋白质(O00187)包括六个功能区(domain)，本实施案例中选取功能区4(CPP1)，功能区5(CPP2)和功能区6(SP)在大肠杆菌进行重组表达。所表达的蛋白质以包涵体形式存在，为了得到有活性的蛋白质，包涵体被复性，然后通过蛋白质片段C末端His进行纯化。获得的重组人MASP-2蛋白分别为hMASP2-D456(其氨基酸序列如SEQ ID NO:1所示),hMASP2A-D456(其氨基酸序列如SEQ ID NO:2所示)。
相同的制备方法用于制备食蟹猴(A0A2K5UJY0|，cMASP2-D456，其氨基酸序列如SEQ ID NO:4所示)和小鼠(Q91WP0，mMASP2-D456，其氨基酸序列如SEQ ID NO:3所示) MASP-2。
实施例2鼠抗人MASP-2单克隆抗体的制备
2.1动物免疫
初次免疫时，使用完全弗氏佐剂依照1:1的比例将上述hMASP2A-D456以及PADRE进行乳化，然后通过腹腔注射至6～8周龄雌性SD大鼠，通过腹腔及皮下多位点注射至6～8周龄雌性Balb/c小鼠，50μg hMASP2A-D456/只；之后间隔两周到三周进行加强免疫，每只大鼠腹腔注射50μg抗原加弗氏不完全佐剂，小鼠采用腹腔及皮下多位点注射，每只小鼠腹腔注射50μg抗原加弗氏不完全佐剂。共计免疫3次，最后一次免疫两周后取动物尾血，测定血清抗人MASP-2抗体滴度。待融合的动物经腹腔注射不加弗氏佐剂的50μg蛋白冲击免疫后，第三天取脾脏细胞。
2.2脾细胞融合
大/小鼠安乐死后，解剖、取脾、研磨并收集细胞，1500rpm，5min离心，收集细胞，用5～10毫升红细胞裂解液悬浮细胞，4℃放置10分钟，用DMEM+10％FBS终止反应，计数。离心后用40ml DMEM悬浮细胞，静置2～3min后转移上清到另外一个50毫升离心管中。收集SP2/0细胞，按SP2/0:脾细胞＝1:2的比例混合，离心，充分吸取上清后，拍松，混合细胞沉淀，用DMEM洗涤混合细胞两次，依照常规方法进行PEG融合。融合后，用DMEM培养基洗涤细胞并将其重悬于DMEM+10％FBS+1×HAT的筛选培养基中。将融合后的细胞加入96孔细胞培养板内，置37℃、湿度75％、5％CO 2培养箱内培养9～10天。
2.3鼠单克隆抗体的筛选
(1)ELISA鉴定杂交瘤单克隆培养上清结合hMASP2-D456功能
用碳酸盐缓冲液稀释hMASP2-D456至1.0μg/mL，加至高结合透明聚苯乙烯96孔板(Nunc)内，100μL/孔，置4℃包被过夜。第二天将ELISA板在自动洗板机上用洗涤缓冲液(PBS+0.05％吐温20(sigma))洗涤二次。每孔加入300μL封闭缓冲液(PBS+0.05％吐温20(sigma)+1％BSA)，置室温封闭1小时。然后在自动洗板机上用洗涤缓冲液洗涤二次，将1000μL杂交瘤上清转移至ELISA板各孔内，置室温孵育1小时，然后依照上述方法洗板3次。每孔加入100μL用封闭缓冲液中1:5000稀释的山羊抗小鼠HRP(Sigma，货号：M4280)；兔抗大鼠Fc-HRP(Sigma，货号：A5795)。置室温孵育1小时，然后依照上述方法洗板3次。加入TMB底物液，100μL/孔，然后每孔加入50μL 1.0M盐酸终止液终止反应，在Thermo Multiscan FC上在450nm处读板。通过结合实验，筛选到杂交瘤克隆大鼠：50A6；小鼠47A1；采用无血清培养及常规抗 体纯化的方法制备杂交瘤单克隆抗体进行抗体功能确认。
实施例3鼠抗人MASP-2单克隆抗体的鉴定
3.1 ELISA鉴定鼠抗人MASP-2单克隆抗体结合人MASP-2功能
用碳酸盐缓冲液(CBS)稀释hMASP2-D456至1.0μg/mL及0.5μg/mL，加至高结合透明聚苯乙烯96孔板(Nunc)内，100μL/孔，置4℃包被过夜。第二天将ELISA板在自动洗板机上用洗涤缓冲液(PBS+0.05％吐温20(sigma))洗涤二次。每孔加入300μL封闭缓冲液(PBS+0.05％吐温20(sigma)+1％BSA)，置室温封闭1小时。然后在自动洗板机上用洗涤缓冲液洗涤二次，用封闭缓冲液将单克隆抗体稀释至，8个梯度。依次加至ELISA板各孔内，置室温孵育1小时，然后依照上述方法洗板3次。每孔加入100μL用封闭缓冲液1:5000稀释的山羊抗小鼠HRP(Sigma，货号：M4280)；兔抗大鼠Fc-HRP(Sigma，货号：A5795)。置室温孵育1小时，然后依照上述方法洗板3次。加入TMB底物液，100μL/孔，然后每孔加入50μL1.0M盐酸终止液终止反应。在Thermo Multiscan FC上在450nm处读板。利用Graphpad作图并计算EC50。结果如图1所示，50A6抗体可以结合人MASP-2；如图2所示，47A1抗体可以结合人MASP-2，本申请鼠抗人MASP-2单克隆抗体均能结合人MASP-2。
3.2 ELISA鉴定鼠抗人MASP-2单克隆抗体交叉结合鼠mMASP2-D456，猴cMASP2-D456功能
用碳酸盐缓冲液稀释鼠mMASP2-D456，食蟹猴cMASP2-D456至1.0μg/mL及0.5μg/mL，加至高结合透明聚苯乙烯96孔板(Nunc)内，100μL/孔，置4℃包被过夜。第二天将ELISA板在自动洗板机上用洗涤缓冲液(PBS+0.05％吐温20(sigma))洗涤二次。每孔加入300μL封闭缓冲液(PBS+0.05％吐温20(sigma)+1％BSA)，置室温封闭1小时。然后在自动洗板机上用洗涤缓冲液洗涤二次，用封闭缓冲液将单克隆抗体50A6稀释至15.0μg/mL，11个梯度。依次加至ELISA板各孔内，置室温孵育1小时，然后依照上述方法洗板3次。每孔加入100μL用封闭缓冲液中1:5000稀释的山羊抗小鼠HRP(Sigma，货号：M4280)；兔抗大鼠Fc-HRP(Sigma，货号：A5795)。置室温孵育1小时，然后依照上述方法洗板3次。加入TMB底物液，100μL/孔，然后每孔加入50μL1.0M盐酸终止液终止反应。在Thermo Multiscan FC上在450nm处读板。利用Graphpad作图并计算EC50。50A6结合猴cMASP2-D456的结果如图3A所示，50A6结合鼠mMASP2-D456的结果如图3B所示。47A1结合猴cMASP2-D456的结果如图4A所示，47A1结合鼠mMASP2-D456的结果如图4B所示。
通过结合、阻断等功能鉴定，选择50A6作为大鼠源候选抗体并进行人源化，选择47A1 作为小鼠源候选抗体并进行人源化。
实施例4杂交瘤单克隆抗体的人源化
4.1通过序列比对鼠源抗体50A6挑选最同源的人种系(Germline)序列(数据来源：IMGT)作为人源化设计框架(轻链以GKV6-21*02，IGKJ2*01为框架；重链以IGHV3-7*01，IGH4*01为框架)；鼠源抗体47A1分别以IGKV1-16*01,IGKJ4*01和IGHV4-30-4*01，IGHJ4*01为轻重链框架。抗体轻重链可变区进行Chothia编号(参见Chothia&Lesk,1987)定义抗体CDR区：CDRL1(L24-L34，即表示VL的第24-34位氨基酸)，CDRL2(L50-L56)，CDRL3(L89-L97)，CDRH1(H26-H32，即表示VH的第26-32位氨基酸)，CDRH2(H52-H56)，CDRH3(H95-H97)，根据序列比对和可变区结构信息对抗体轻重链可变区氨基酸进行人源化突变。
4.2设计表达载体，基因合成，哺乳细胞表达纯化重组抗体，比较人源化改造后抗体活性，理化性质的差异，进行1-2轮人源化优化。
4.3以上述种系(Germline)抗体为框架进行CDR移植，先获得以下的嵌合抗体的可变区氨基酸序列：50A6的轻链可变区序列(其氨基酸序列如SEQ ID NO:5所示)；50A6的重链可变区序列(其氨基酸序列如SEQ ID NO:13所示)。
47A1的轻链可变区序列(其氨基酸序列如SEQ ID NO:31所示)；47A1的重链可变区序列(其氨基酸序列如SEQ ID NO:39所示)。
再在上述嵌合抗体的可变区氨基酸序列的基础上进行氨基酸突变，获得以下的人源化优化氨基酸序列：JYB1931A63的轻链可变区序列(其氨基酸序列如SEQ ID NO:21所示)，JYB1931A63的重链可变区序列(其氨基酸序列如SEQ ID NO:26所示)。JYB1931A13的轻链可变区序列(其氨基酸序列如SEQ ID NO:47所示)；JYB1931A13的重链可变区序列(其氨基酸序列如SEQ ID NO:52所示)。
4.4抗体表达和纯化
设计上述人源化优化氨基酸序列后，将其分别进行密码子优化，并合成对应的DNA基因片段(Genscript)，扩增合成的所述基因片段到表达载体pcDNA3.4(Life Technologies)。表达质粒扩增和质粒抽提后双质粒共转ExpiCHO细胞(ThermoFisher Scientific，A29133)，根据供应商ExpiCHO表达系统方法进行抗体瞬转表达，过程如下：在培养总体积25mL培养基中，36.5℃，8％二氧化碳浓度下培养ExpiCHO细胞至密度为6×10 6/mL，使用ExpiFectamine转染试剂化转各10μg抗体轻重链表达质粒到细胞；转染一天后，各取150μL和4mL ExpiCHO增 强剂和ExpiCHO辅料添加到培养细胞中，继续培养至9天，4℃，3500转离心取上清。混合AmMagTM Protein A磁珠(Genscript，L00695)和抗体表达上清，室温孵育2小时，PBS洗涤两次弃上清，加入适量洗脱缓冲液蛋白G或A SefinoseTMElution缓冲液(Sangon,C600481)，充分混匀后置于试管架上静止孵育5分钟，孵育期间重悬磁珠2～3次，重复洗脱2次，洗脱后，立即加入适量中和液1M Tris-HCl，pH7.5(Sangon,B548124)中和备用。
实施例5抗MASP-2鼠源抗体阻断人补体系统凝集素通路
5.1采用
Figure PCTCN2022097268-appb-000001
补体系统凝集素通路(Svar Life Science AB，货号：AS 1327)试剂盒检测抗体对人血清补体信号通路阻断。用三种稀释液稀释人血清和抗体50A6和47A1：其中人血清稀释方案为CP:80ul/4ml、MP:80ul/4ml和AP:444.7/4ml，抗体以500nM起，10倍稀释。
5.2实验步骤
首先将稀释后的样品和血清加入板中，各50μL，37℃孵育1h；2)用洗涤缓冲液洗板三次；3)每孔加入100μL偶联物室温放置30min；4)洗板：用洗涤缓冲液洗板三次；5)每孔加入100μL底物溶液室温放置30min；6)每孔加入100μL 5mM EDTA终止显色；7)终止显色后多谱酶联阅读仪(酶标仪)读取450nm的吸光值。
5.3数据处理：数据用GraphPad Prism软件进行处理。如图5所示，本申请的抗体可以阻断人补体系统凝集素通路。
实施例6抗MASP-2抗体的亲和力测定
6.1样品稀释
采用Octet RED96e(Fortebio)测定候选抗体JYB1931A63和JYB1931A13与人hMASP2-D456(KACTUS，批号：080203)、猴cMASP2-D456(KACTUS，批号：030301)和鼠mMASP2-D456(KACTUS，批号：030401)的亲和力，抗原及抗体均用1xPBST(1xPBS：生工，B548117-0500；0.02％吐温20：sigma-aldrich，P1379)稀释，抗原使用浓度为30nM，抗体使用浓度为5μg/mL。
6.2样品上机检测(Octet Data Acquisition 11.1.0.11)
首先，将样品加入96孔板(Greiner bio-one，655209)，体系为200μL/孔。然后设置软件参数，板温设定为30℃，收集标准动力学信号的频率为5.0HZ。接着，用1xPBST预湿AHC传感器(Fortébio,货号：18-0015)10分钟，然后上机检测。每个循环包含以下步骤：1)浸 入缓冲液60s；2)检测抗原是否与传感器有非特异性结合；3)10mM pH 1.7的甘氨酸溶液再生；4)浸入缓冲液60s；5)抗体固化在传感器上，时间为60s；6)传感器浸入缓冲液180s；7)抗原与抗体结合，时间180s；8)抗原抗体的解离，时间5分钟；9)传感器再生。6.3数据分析
采用Fortebio的Data Analysis 12.0软件，对抗原-抗体以1:1的结合方式，测定结合速率(Ka)和解离速率(Kd)，以此计算抗体的平衡解离常数(K D)。结果如表1所示。
表1抗体与人hMASP2-D456、猴cMASP2-D456和鼠mMASP2-D456的亲和力
Figure PCTCN2022097268-appb-000002
实施例7抗MASP-2人源化抗体阻断补体系统凝集素通路
采用
Figure PCTCN2022097268-appb-000003
补体系统凝集素通路(Svar Life Science AB，货号：AS 1327)试剂盒检测抗体对人血清和猴血清的补体信号通路阻断。用三种稀释液稀释人血清和猴血清：其中人血清稀释方案为CP:80ul/4ml、MP:80ul/4ml和AP:444.7/4ml；猴血清稀释方案为CP:2/100、MP:2/100和AP:11.1/100。样品分别用三种稀释液稀释：500nM起，10倍稀释。实验步骤和数据处理同实施例5。如图6所示，本申请的人源化抗体可以阻断补体系统凝集素通路。
实施例8抗体物理化学性质检测
8.1 SEC-HPLC纯度分析
(1)将样品稀释至1mg/mL，混匀，12000rpm离心5min，取上清转至样品瓶，放入HPLC样品盘。设置色谱条件如表2所示。
表2色谱条件
Figure PCTCN2022097268-appb-000004
(2)色谱柱采用流动相(200mM磷酸盐缓冲液，pH6.8)平衡后，进样分析，用色谱软件进行数据分析，峰面积归一化法计算各个峰的峰面积百分比。
8.2 HIC-HPLC分析
(1)将样品稀释至1mg/ml，离心取上清待测。设置色谱条件如下(表3)：
表3色谱条件
Figure PCTCN2022097268-appb-000005
(2)用流动相A(50mM磷酸盐缓冲液/1M硫酸铵，pH 7.0)和流动相B(50mM磷酸盐缓冲液，pH 7.0)进行梯度洗脱，记录主峰保留时间。
8.3熔解温度(Tm)值分析
将供试品用样品缓冲液稀释至1mg/mL，然后按照Protein Thermal Shift TM Starter Kit说明书，取供试品溶液13μL加入至PCR管内，加入5μL Protein Thermal shift  TM Buffer，加入2μL10×染色液，使反应体积为20μL，混匀后，12000rpm离心5min以去除气泡。将检测样品置于PCR仪内，进行样品分析，记录样品的Tm值。
8.4 iCIEF分析
取样品溶液加入到已经充分混匀的以下体系中：1％的甲基纤维素(MC)70μl，尿素5M80μl，两性电解质Pharmalyte pH 3-10 8μl，pI marker 5.5和9.5各1μl。补加适当体积超纯水至200μl，混匀。离心取上清进样分析。分析结束后，将结果文件导入ChromPerfect软件进行图谱积分处理并计算各峰的等电点以及各峰百分比，如表4所示。
表4理化分析结果
Figure PCTCN2022097268-appb-000006
Figure PCTCN2022097268-appb-000007
实施例9抗体分子在人源化FcRn小鼠模型中药代动力学(PK)研究
以人源化FcRn小鼠为受试动物，分别研究两个待试药物阳性对照(Narsoplimab),JYB1931A63单次腹腔给药后的药代动力学指标。所有动物实验方案获得了IACUC审阅和批准。hFcRn小鼠采购自北京百奥赛图，雄性，6～8周龄，体重23～26g，饲养于SPF级动物房，标准颗粒饲料喂养，自由进食及饮水，室温18～24℃，相对湿度40％～50％，每日12h昼夜交替。实验动物共16只，随机分为四组，每组四只动物，腹腔单次给药，给药剂量为10mg/kg，给药体积为10mL/kg。采血时间点为给药前，给药后2h，6h，24h(第1天)，第2天，第3天，第4天，第7天，第10天，第14天，第21天，第28天，第35天和第42天。通过眼眶采集全血60μL/只至EP管中，室温静置30min，然后离心(2000g，4℃，5min)分离血清，每个样品分装为2份(检测管和备份管)，10μL/管，置于-80℃保存。
利用Elisa间接法检测，分析四种药下不同时间点药代动力学。包被抗原为重组人MASP-2蛋白，1μg/mL，100μL/孔，4℃，过夜。洗板后用200μL/孔封闭液4℃过夜。血清样本加样，50μL/孔，37℃，1h。检测抗体为小鼠单克隆抗体[H2]抗人IgG F(ab)'2(HRP)(abcam，ab87422，GR3246767-11)+Streptavidin-peroxidase(Sigma，lot:SLCB5784)，100μl/孔，37℃，0.5h。TMB显色液(KPL，货号：52-00-03)显色，酶标仪(Molecular Devices，SpectraMax M3)读值OD450。根据标准曲线获得药物浓度，PK Solver非房室进行数据处理获得PK参数。血药浓度曲线如图7所示。
给药阳性对照后，半衰期(t 1/2)＝4.93天，峰值时间点(Tmax)为0.45天最高浓度(Cmax)为9393ng/ml；给药JYB1931A63后半衰期(t 1/2)＝18.6天，峰值时间点(Tmax)为4.53天，最高浓度(Cmax)为86934ng/ml。
前述详细说明是以解释和举例的方式提供的，并非要限制所附权利要求的范围。目前本申请所列举的实施方式的多种变化对本领域普通技术人员来说是显而易见的，且保留在所附的权利要求和其等同方案的范围内。

Claims (102)

  1. 分离的抗原结合蛋白，其具有下述性质中的一种或多种：
    a)在Octet检测中，以约2E-09M或以下的K D值与人MASP-2蛋白特异性结合；
    b)在Octet检测中，以约2E-09M或以下的K D值与猴MASP-2蛋白特异性结合；
    c)能够特异性阻断人补体系统凝集素通路而不影响补体经典路径和旁路途径。
  2. 根据权利要求1所述的分离的抗原结合蛋白，其包含HCDR3，所述HCDR3包含SEQ ID NO:19所示的氨基酸序列。
  3. 根据权利要求1-2中任一项所述的分离的抗原结合蛋白，其包含HCDR2，所述HCDR2包含SEQ ID NO:17所示的氨基酸序列。
  4. 根据权利要求1-3中任一项所述的分离的抗原结合蛋白，其包含HCDR1，所述HCDR1包含SEQ ID NO:15所示的氨基酸序列。
  5. 根据权利要求1-4中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO：67所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
  6. 根据权利要求1-5中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO：13和SEQ ID NO：26中任一项所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
  7. 根据权利要求1-6中任一项所述的分离的抗原结合蛋白，其包括重链可变区VH，所述VH包含所述HCDR1、HCDR2和HCDR3，所述HCDR3包含SEQ ID NO:19所示的氨基酸序列；所述HCDR2包含SEQ ID NO:17所示的氨基酸序列；以及所述HCDR1包含SEQ ID NO:15所示的氨基酸序列。
  8. 根据权利要求1-7中任一项所述的分离的抗原结合蛋白，其包含H-FR1，所述H-FR1的C末端与所述HCDR1的N末端直接或间接地相连，且所述H-FR1包含SEQ ID NO：62所示的氨基酸序列。
  9. 根据权利要求8所述的分离的抗原结合蛋白，其中所述H-FR1包含SEQ ID NO:14和SEQ ID NO:27中任一项所示的氨基酸序列。
  10. 根据权利要求1-9中任一项所述的分离的抗原结合蛋白，其包含H-FR2，所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO：63所示的氨基酸序列。
  11. 根据权利要求10所述的分离的抗原结合蛋白，其中所述H-FR2包含SEQ ID NO:16和SEQ ID NO:28中任一项所示的氨基酸序列。
  12. 根据权利要求1-11中任一项所述的分离的抗原结合蛋白，其包含H-FR3，所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO：64所示的氨基酸 序列。
  13. 根据权利要求12所述的分离的抗原结合蛋白，其中所述H-FR3包含SEQ ID NO:18和SEQ ID NO:29中任一项所示的氨基酸序列。
  14. 根据权利要求1-13中任一项所述的分离的抗原结合蛋白，其包含H-FR4，所述H-FR4的N末端与所述HCDR3的C末端直接或间接地相连，且所述H-FR4包含SEQ ID NO：65所示的氨基酸序列。
  15. 根据权利要求14所述的分离的抗原结合蛋白，其中所述H-FR4包含SEQ ID NO:20和SEQ ID NO:30中任一项所示的氨基酸序列。
  16. 根据权利要求1-15中任一项所述的分离的抗原结合蛋白，其包含H-FR1，H-FR2，H-FR3和H-FR4，所述H-FR1包含SEQ ID NO：62所示的氨基酸序列；所述H-FR2包含SEQ ID NO：63所示的氨基酸序列；所述H-FR3包含SEQ ID NO：64所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO：65所示的氨基酸序列。
  17. 根据权利要求16所述的分离的抗原结合蛋白，其中所述H-FR1包含SEQ ID NO:14和SEQ ID NO:27中任一项所示的氨基酸序列；所述H-FR2包含SEQ ID NO:16和SEQ ID NO:28中任一项所示的氨基酸序列；所述H-FR3包含SEQ ID NO:18和SEQ ID NO:29中任一项所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO:20和SEQ ID NO:30中任一项所示的氨基酸序列。
  18. 根据权利要求16-17中任一项所述的分离的抗原结合蛋白，其中所述H-FR1、H-FR2、H-FR3和H-FR4包含选自下述任意一组的氨基酸序列：
    a)H-FR1：SEQ ID NO:14，H-FR2：SEQ ID NO:16，H-FR3：SEQ ID NO:18和H-FR4：SEQ ID NO:20；
    b)H-FR1：SEQ ID NO:27，H-FR2：SEQ ID NO:28，H-FR3：SEQ ID NO:29和H-FR4：SEQ ID NO:30。
  19. 根据权利要求1-18中任一项所述的分离的抗原结合蛋白，其包含重链可变区VH，所述VH包含SEQ ID NO：67所示的氨基酸序列。
  20. 根据权利要求19所述的分离的抗原结合蛋白，其中所述VH包含SEQ ID NO:13和SEQ ID NO:26中任一项所示的氨基酸序列。
  21. 根据权利要求1-20中任一项所述的分离的抗原结合蛋白，其包含LCDR3，所述LCDR3包含SEQ ID NO：11所示的氨基酸序列。
  22. 根据权利要求1-21中任一项所述的分离的抗原结合蛋白，其包含LCDR2，所述LCDR2 包含SEQ ID NO：9所示的氨基酸序列。
  23. 根据权利要求1-22中任一项所述的分离的抗原结合蛋白，其包含LCDR1，所述LCDR1包含SEQ ID NO：7所示的氨基酸序列。
  24. 根据权利要求1-23中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO：66所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
  25. 根据权利要求1-24中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO：5和21中任一项所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
  26. 根据权利要求1-25中任一项所述的分离的抗原结合蛋白，其包含轻链可变区VL，所述VL包含所述LCDR1、LCDR2和LCDR3，所述LCDR3包含SEQ ID NO：11所示的氨基酸序列；所述LCDR2包含SEQ ID NO：9所示的氨基酸序列；以及所述LCDR1包含SEQ ID NO：7所示的氨基酸序列。
  27. 根据权利要求1-26中任一项所述的分离的抗原结合蛋白，其包含L-FR1，所述L-FR1的C末端与所述LCDR1的N末端直接或间接地相连，且所述L-FR1包含SEQ ID NO：58所示的氨基酸序列。
  28. 根据权利要求27所述的分离的抗原结合蛋白，其中所述L-FR1包含SEQ ID NO:6和SEQ ID NO:22中任一项所示的氨基酸序列。
  29. 根据权利要求1-28中任一项所述的分离的抗原结合蛋白，其包含L-FR2，所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO：59所示的氨基酸序列。
  30. 根据权利要求29所述的分离的抗原结合蛋白，其中所述L-FR2包含SEQ ID NO:8和SEQ ID NO:23中任一项所示的氨基酸序列。
  31. 根据权利要求1-30中任一项所述的分离的抗原结合蛋白，其包含L-FR3，所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO：60所示的氨基酸序列。
  32. 根据权利要求31所述的分离的抗原结合蛋白，其中所述L-FR3包含SEQ ID NO:10和SEQ ID NO:24中任一项所示的氨基酸序列。
  33. 根据权利要求1-32中任一项所述的分离的抗原结合蛋白，其包含L-FR4，所述L-FR4的N末端与所述LCDR3的C末端直接或间接地相连，且所述L-FR4包含SEQ ID NO：61所示的氨基酸序列。
  34. 根据权利要求33所述的分离的抗原结合蛋白，其中所述L-FR4包含SEQ ID NO:12和SEQ ID NO:25中任一项所示的氨基酸序列。
  35. 根据权利要求1-34中任一项所述的分离的抗原结合蛋白，其包含L-FR1，L-FR2，L-FR3和L-FR4，所述L-FR1包含SEQ ID NO：58所示的氨基酸序列；所述L-FR2包含SEQ ID NO：59所示的氨基酸序列；所述L-FR3包含SEQ ID NO：60所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO：61所示的氨基酸序列。
  36. 根据权利要求35所述的分离的抗原结合蛋白，其中所述L-FR1包含SEQ ID NO:6和SEQ ID NO:22中任一项所示的氨基酸序列；所述L-FR2包含SEQ ID NO:8和SEQ ID NO:23中任一项所示的氨基酸序列；所述L-FR3包含SEQ ID NO:10和SEQ ID NO:24中任一项所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO:12和SEQ ID NO:25中任一项所示的氨基酸序列。
  37. 根据权利要求35-36中任一项所述的分离的抗原结合蛋白，其中所述L-FR1、L-FR2、L-FR3和L-FR4包含选自下述任意一组的氨基酸序列：
    a)L-FR1：SEQ ID NO:6，L-FR2：SEQ ID NO:8，L-FR3：SEQ ID NO:10和L-FR4：SEQ ID NO:12；
    b)L-FR1：SEQ ID NO:22，L-FR2：SEQ ID NO:23，L-FR3：SEQ ID NO:24和L-FR4：SEQ ID NO:25。
  38. 根据权利要求1-37中任一项所述的分离的抗原结合蛋白，其中所述VL包含SEQ ID NO：66所示的氨基酸序列。
  39. 根据权利要求38所述的分离的抗原结合蛋白，其中所述VL包含SEQ ID NO:5和SEQ ID NO:21中任一项所示的氨基酸序列。
  40. 根据权利要求1-39中任一项所述的分离的抗原结合蛋白，其包括VH和VL，其中所述VH和VL包含选自下述任意一组的氨基酸序列：
    a)VH：SEQ ID NO:13和VL：SEQ ID NO:5；
    b)VH：SEQ ID NO:26和VL：SEQ ID NO:21。
  41. 根据权利要求1所述的分离的抗原结合蛋白，其包含HCDR3，所述HCDR3包含SEQ ID NO：45所示的氨基酸序列。
  42. 根据权利要求1和41中任一项所述的分离的抗原结合蛋白，其包含HCDR2，所述HCDR2包含SEQ ID NO：43所示的氨基酸序列。
  43. 根据权利要求1和41-42中任一项所述的分离的抗原结合蛋白，其包含HCDR1，所述HCDR1包含SEQ ID NO：41所示的氨基酸序列。
  44. 根据权利要求1和41-43中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO：77 所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
  45. 根据权利要求1和41-44中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO:39和SEQ ID NO:52所示的重链可变区VH的HCDR1、HCDR2和HCDR3。
  46. 根据权利要求1和41-45中任一项所述的分离的抗原结合蛋白，其包括重链可变区VH，所述VH包含所述HCDR1、HCDR2和HCDR3，所述HCDR3包含SEQ ID NO：45所示的氨基酸序列；所述HCDR2包含SEQ ID NO：43所示的氨基酸序列；以及所述HCDR1包含SEQ ID NO：41所示的氨基酸序列。
  47. 根据权利要求1和41-46中任一项所述的分离的抗原结合蛋白，其包含H-FR1，所述H-FR1的C末端与所述HCDR1的N末端直接或间接地相连，且所述H-FR1包含SEQ ID NO：72所示的氨基酸序列。
  48. 根据权利要求47所述的分离的抗原结合蛋白，其中所述H-FR1包含SEQ ID NO:40和SEQ ID NO:53中任一项所示的氨基酸序列。
  49. 根据权利要求1和41-48中任一项所述的分离的抗原结合蛋白，其包含H-FR2，所述H-FR2位于所述HCDR1与所述HCDR2之间，且所述H-FR2包含SEQ ID NO：73所示的氨基酸序列。
  50. 根据权利要求49所述的分离的抗原结合蛋白，其中所述H-FR2包含SEQ ID NO:42和SEQ ID NO:54中任一项所示的氨基酸序列。
  51. 根据权利要求1和41-50中任一项所述的分离的抗原结合蛋白，其包含H-FR3，所述H-FR3位于所述HCDR2与所述HCDR3之间，且所述H-FR3包含SEQ ID NO：74所示的氨基酸序列。
  52. 根据权利要求51所述的分离的抗原结合蛋白，其中所述H-FR3包含SEQ ID NO:44和SEQ ID NO:55中任一项所示的氨基酸序列。
  53. 根据权利要求1和41-52中任一项所述的分离的抗原结合蛋白，其包含H-FR4，所述H-FR4的N末端与所述HCDR3的C末端直接或间接地相连，且所述H-FR4包含SEQ ID NO：75所示的氨基酸序列。
  54. 根据权利要求53所述的分离的抗原结合蛋白，其中所述H-FR4包含SEQ ID NO:46和SEQ ID NO:30中任一项所示的氨基酸序列。
  55. 根据权利要求1和41-54中任一项所述的分离的抗原结合蛋白，其包含H-FR1，H-FR2，H-FR3和H-FR4，所述H-FR1包含SEQ ID NO：72所示的氨基酸序列；所述H-FR2包含SEQ ID NO：73所示的氨基酸序列；所述H-FR3包含SEQ ID NO：74所示的氨基酸 序列；以及所述H-FR4包含SEQ ID NO：75所示的氨基酸序列。
  56. 根据权利要求55所述的分离的抗原结合蛋白，其中所述H-FR1包含SEQ ID NO:40和SEQ ID NO:53中任一项所示的氨基酸序列；所述H-FR2包含SEQ ID NO:42和SEQ ID NO:54中任一项所示的氨基酸序列；所述H-FR3包含SEQ ID NO:44和SEQ ID NO:55中任一项所示的氨基酸序列；以及所述H-FR4包含SEQ ID NO:46和SEQ ID NO:30中任一项所示的氨基酸序列。
  57. 根据权利要求55-56中任一项所述的分离的抗原结合蛋白，其中所述H-FR1、H-FR2、H-FR3和H-FR4包含选自下述任意一组的氨基酸序列：
    a)H-FR1：SEQ ID NO:40，H-FR2：SEQ ID NO:42，H-FR3：SEQ ID NO:44和H-FR4：SEQ ID NO:46；
    b)H-FR1：SEQ ID NO:53，H-FR2：SEQ ID NO:54，H-FR3：SEQ ID NO:55和H-FR4：SEQ ID NO:30。
  58. 根据权利要求1和41-57中任一项所述的分离的抗原结合蛋白，其包含重链可变区VH，所述VH包含SEQ ID NO：77所示的氨基酸序列。
  59. 根据权利要求58所述的分离的抗原结合蛋白，其中所述VH包含SEQ ID NO:39和SEQ ID NO:52中任一项所示的氨基酸序列。
  60. 根据权利要求1和41-59中任一项所述的分离的抗原结合蛋白，其包含LCDR3，所述LCDR3包含SEQ ID NO：37所示的氨基酸序列。
  61. 根据权利要求1和41-60中任一项所述的分离的抗原结合蛋白，其包含LCDR2，所述LCDR2包含SEQ ID NO：35所示的氨基酸序列。
  62. 根据权利要求1和41-61中任一项所述的分离的抗原结合蛋白，其包含LCDR1，所述LCDR1包含SEQ ID NO：33所示的氨基酸序列。
  63. 根据权利要求1和41-62中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO：76所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
  64. 根据权利要求1和41-63中任一项所述的分离的抗原结合蛋白，其包含SEQ ID NO:31和SEQ ID NO:47所示的轻链可变区VL的LCDR1、LCDR2和LCDR3。
  65. 根据权利要求1和41-64中任一项所述的分离的抗原结合蛋白，其包含轻链可变区VL，所述VL包含所述LCDR1、LCDR2和LCDR3，所述LCDR3包含SEQ ID NO：37所示的氨基酸序列；所述LCDR2包含SEQ ID NO：35所示的氨基酸序列；以及所述LCDR1包含SEQ ID NO：33所示的氨基酸序列。
  66. 根据权利要求1和41-65中任一项所述的分离的抗原结合蛋白，其包含L-FR1，所述L- FR1的C末端与所述LCDR1的N末端直接或间接地相连，且所述L-FR1包含SEQ ID NO：68所示的氨基酸序列。
  67. 根据权利要求66所述的分离的抗原结合蛋白，其中所述L-FR1包含SEQ ID NO:32和SEQ ID NO:48中任一项所示的氨基酸序列。
  68. 根据权利要求1和41-67中任一项所述的分离的抗原结合蛋白，其包含L-FR2，所述L-FR2位于所述LCDR1与所述LCDR2之间，且所述L-FR2包含SEQ ID NO：69所示的氨基酸序列。
  69. 根据权利要求68所述的分离的抗原结合蛋白，其中所述L-FR2包含SEQ ID NO:34和SEQ ID NO:49中任一项所示的氨基酸序列。
  70. 根据权利要求1和41-69中任一项所述的分离的抗原结合蛋白，其包含L-FR3，所述L-FR3位于所述LCDR2与所述LCDR3之间，且所述L-FR3包含SEQ ID NO：70所示的氨基酸序列。
  71. 根据权利要求70所述的分离的抗原结合蛋白，其中所述L-FR3包含SEQ ID NO:36和SEQ ID NO:50中任一项所示的氨基酸序列。
  72. 根据权利要求1和41-71中任一项所述的分离的抗原结合蛋白，其包含L-FR4，所述L-FR4的N末端与所述LCDR3的C末端直接或间接地相连，且所述L-FR4包含SEQ ID NO：71所示的氨基酸序列。
  73. 根据权利要求72所述的分离的抗原结合蛋白，其中所述L-FR4包含SEQ ID NO:38和SEQ ID NO:51中任一项所示的氨基酸序列。
  74. 根据权利要求1和41-73中任一项所述的分离的抗原结合蛋白，其包含L-FR1，L-FR2，L-FR3和L-FR4，所述L-FR1包含SEQ ID NO：68所示的氨基酸序列；所述L-FR2包含SEQ ID NO：69所示的氨基酸序列；所述L-FR3包含SEQ ID NO：70所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO：71所示的氨基酸序列。
  75. 根据权利要求74所述的分离的抗原结合蛋白，其中所述L-FR1包含SEQ ID NO:32和SEQ ID NO:48中任一项所示的氨基酸序列；所述L-FR2包含SEQ ID NO:34和SEQ ID NO:49中任一项所示的氨基酸序列；所述L-FR3包含SEQ ID NO:36和SEQ ID NO:50中任一项所示的氨基酸序列；以及所述L-FR4包含SEQ ID NO:38和SEQ ID NO:51中任一项所示的氨基酸序列。
  76. 根据权利要求74-75中任一项所述的分离的抗原结合蛋白，其中所述L-FR1、L-FR2、L-FR3和L-FR4包含选自下述任意一组的氨基酸序列：
    a)L-FR1：SEQ ID NO:32，L-FR2：SEQ ID NO:34，L-FR3：SEQ ID NO:36和L-FR4：SEQ ID NO:38；
    b)L-FR1：SEQ ID NO:48，L-FR2：SEQ ID NO:49，L-FR3：SEQ ID NO:50和L-FR4：SEQ ID NO:51。
  77. 根据权利要求1和41-76中任一项所述的分离的抗原结合蛋白，其包括VL，所述VL包含SEQ ID NO：76所示的氨基酸序列。
  78. 根据权利要求77所述的分离的抗原结合蛋白，其中所述VL包含SEQ ID NO：31和47中任一项所示的氨基酸序列。
  79. 根据权利要求1和41-78中任一项所述的分离的抗原结合蛋白，其包括VH和VL，所述VH和VL包含选自下述任意一组的氨基酸序列：
    a)VH：SEQ ID NO:39和VL：SEQ ID NO:31；
    b)VH：SEQ ID NO:52和VL：SEQ ID NO:47。
  80. 根据权利要求1-79中任一项所述的分离的抗原结合蛋白，其包含重链恒定区，且所述重链恒定区包括源自IgG的恒定区或源自IgY的恒定区。
  81. 根据权利要求80所述的分离的抗体结合蛋白，其中所述重链恒定区包括源自IgG的恒定区。
  82. 根据权利要求80-81中任一项所述的分离的抗原结合蛋白，其中所述重链恒定区包括源自IgG1、IgG2、IgG3或IgG4的恒定区。
  83. 根据权利要求80-82中任一项所述的分离的抗原结合蛋白，其中所述重链恒定区包含SEQ ID NO：56所示的氨基酸序列。
  84. 根据权利要求1-83中任一项所述的分离的抗原结合蛋白，其包含轻链恒定区，且所述轻链恒定区包括源自Igκ的恒定区或源自Igλ的恒定区。
  85. 根据权利要求84所述的分离的抗原结合蛋白，其中所述轻链恒定区包括源自人Igκ的恒定区。
  86. 根据权利要求84-85中任一项所述的分离的抗原结合蛋白，其中所述轻链恒定区包含SEQ ID NO：57所示的氨基酸序列。
  87. 根据权利要求1-86中任一项所述的分离的抗原结合蛋白，其包括抗体或其抗原结合片段。
  88. 根据权利要求87所述的分离的抗原结合蛋白，其中所述抗原结合片段选自下组：Fab，Fab’，F(ab)2，Fv片段，F(ab’)2，scFv，di-scFv，VHH和/或dAb。
  89. 根据权利要求87-88中任一项所述的分离的抗原结合蛋白，其中所述抗体选自下组：单 克隆抗体、单链抗体、嵌合抗体、人源化抗体和全人源抗体。
  90. 多肽，其包含权利要求1-89中任一项所述的分离的抗原结合蛋白。
  91. 免疫缀合物，其包含权利要求1-89中任一项所述的分离的抗原结合蛋白或权利要求90所述的多肽。
  92. 分离的核酸分子，其编码权利要求1-89中任一项所述的分离的抗原结合蛋白，或者权利要求90所述的多肽。
  93. 载体，其包含权利要求92所述的分离的核酸分子。
  94. 细胞，其包含权利要求1-89中任一项所述的分离的抗原结合蛋白，权利要求90所述的多肽，权利要求91所述的免疫缀合物，权利要求92所述的分离的核酸分子和/或权利要求93所述的载体。
  95. 制备权利要求1-89中任一项所述的分离的抗原结合蛋白或权利要求90所述的多肽的方法，所述方法包括再使得权利要求1-89中任一项所述的分离的抗原结合蛋白或权利要求90所述的多肽表达的条件下，培养权利要求94所述的细胞。
  96. 药物组合物，其包含权利要求1-89中任一项所述的分离的抗原结合蛋白，权利要求90所述的多肽，权利要求91所述的免疫缀合物，权利要求92所述的分离的核酸分子，权利要求93所述的载体，权利要求94所述的细胞，和/或药学上可接受的佐剂和/或赋形剂。
  97. 一种用于检测或测定MASP-2的方法，所述方法包括使用权利要求1-89中任一项所述的分离的抗原结合蛋白或权利要求90所述的多肽。
  98. 一种MASP-2的检测试剂盒，其包含权利要求1-89中任一项所述的分离的抗原结合蛋白或权利要求90所述的多肽。
  99. 权利要求1-89中任一项所述的分离的抗原结合蛋白或权利要求90所述的多肽在制备试剂盒中的用途，所述试剂盒用于检测MASP-2的存在和/或含量。
  100. 权利要求1-89中任一项所述的分离的抗原结合蛋白，权利要求90所述的多肽，权利要求91所述的免疫缀合物，权利要求92所述的分离的核酸分子，权利要求93所述的载体，权利要求94所述的细胞和/或权利要求96所述的药物组合物在制备预防和/或治疗疾病或病症的药物中的用途。
  101. 权利要求1-89中任一项所述的分离的抗原结合蛋白，权利要求90所述的多肽，权利要求91所述的免疫缀合物，权利要求92所述的分离的核酸分子，权利要求93所述的载体，权利要求94所述的细胞和/或权利要求96所述的药物组合物，其用于预防、缓解和/或治疗疾病或病症。
  102. 一种预防和/或治疗疾病或病症的方法，其包括向有需要的受试者施用有效量的权利要求1-89中任一项所述的分离的抗原结合蛋白，权利要求90所述的多肽，权利要求91所述的免疫缀合物，权利要求92所述的分离的核酸分子，权利要求93所述的载体，权利要求94所述的细胞和/或权利要求96所述的药物组合物。
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