Anti-MASP-2 antibody and its use

Abstract

K below 2E-09M D specifically binds to human MASP-2 protein with a K value of about 2E-09M or less D An anti-MASP-2 antibody that specifically binds to cynomolgus monkey MASP-2 protein at a specific value and uses thereof. An immunoconjugate containing said anti-MASP-2 antibody, a method for producing said anti-MASP-2 antibody, and uses of said anti-MASP-2 antibody.

Description

Technical Field

[0001] The present application relates to the field of biopharmaceuticals, and specifically to anti-MASP-2 antibodies and their use.

Background Art

[0002] Immunoglobulin A nephropathy (IgAN) (also called Berger's disease) is a mesangial proliferative glomerulonephritis (GN) characterized by diffuse deposition of IgA in the renal mesangium. Currently, for the treatment of IgA nephropathy, the KIDGO guidelines recommend the use of ACEi / ARB to reduce urinary protein and improve kidney function. Also, immunofluorescence studies have shown that local complement C3 activation in glomeruli is associated with poor prognosis, suggesting that IgA nephropathy is related to complement system activation. The complement system includes three pathways: 1) the classical pathway (CP classical pathway), 2) the alternative pathway (AP alternative pathway), and 3) the lectin pathway (LP lectin pathway). Previously, in the medical community, it was generally thought that the prognosis of this disease was good and it did not harm health. However, as the understanding of IgA nephropathy has deepened over time, physicians have discovered that not all IgA nephropathy patients have a good prognosis, and a significant proportion of IgA nephropathy patients have a poor prognosis and progress to uremia 10 to 20 years after onset. If it is IgA nephropathy that is not properly controlled and progresses rapidly, it will impose a great burden on the family and society. However, currently, there are relatively few "weapons" for treating IgA nephropathy. Although the condition of IgA nephropathy patients can be controlled with RAS blockers (sartan drugs and pril drugs) and hormones, a significant proportion of patients have a poor response to these drugs. Kidney damage caused by abnormal activation of the complement is a very important direction in current basic research, and part of the deterioration of IgA nephropathy is caused by abnormal activation of the complement.

[0003] MASP-2 is an effector enzyme of the lectin signaling pathway in the complement system and is one of the suitable targets for preventing abnormal activation of the complement.

[0004] Therefore, there is an urgent need to develop a novel anti-MASP-2 antibody with high affinity and high specificity for MASP-2.

Summary of the Invention

[0005] This application provides an isolated antigen-binding protein having one or more of the following properties. 1) Specifically bind to human MASP-2 protein with a KD value of about 2E-09 M or less in Octet detection, 2) Specifically bind to monkey MASP-2 protein with a KD value of about 2E-09 M or less in Octet detection, 3) Specifically block the human complement lectin pathway without affecting the classical and alternative pathways of the complement

[0006] In some embodiments, the isolated antigen-binding protein comprises an HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19.

[0007] In some embodiments, the isolated antigen-binding protein comprises an HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 17.

[0008] In some embodiments, the isolated antigen-binding protein comprises an HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 15.

[0009] In some embodiments, the isolated antigen-binding protein comprises HCDR1, HCDR2, and HCDR3 of the heavy chain variable region VH shown in SEQ ID NO: 67.

[0010] In some embodiments, the isolated antigen-binding protein comprises HCDR1, HCDR2, and HCDR3 of the heavy-chain variable region VH represented by SEQ ID NO: 13 and SEQ ID NO: 26.

[0011] In some embodiments, the isolated antigen-binding protein comprises a heavy-chain variable region VH comprising the HCDR1, HCDR2, and HCDR3, wherein the HCDR3 comprises the amino acid sequence represented by SEQ ID NO: 19, the HCDR2 comprises the amino acid sequence represented by SEQ ID NO: 17, and the HCDR1 comprises the amino acid sequence represented by SEQ ID NO: 15.

[0012] In some embodiments, the isolated antigen-binding protein comprises an H-FR1 having a C-terminus directly or indirectly linked to the N-terminus of the HCDR1 and comprising the amino acid sequence represented by SEQ ID NO: 62.

[0013] In some embodiments, the H-FR1 comprises the amino acid sequence represented by either SEQ ID NO: 14 or SEQ ID NO: 27.

[0014] In some embodiments, the isolated antigen-binding protein comprises an H-FR2 located between the HCDR1 and the HCDR2 and comprising the amino acid sequence represented by SEQ ID NO: 63.

[0015] In some embodiments, the H-FR2 comprises the amino acid sequence represented by either SEQ ID NO: 16 or SEQ ID NO: 28.

[0016] In some embodiments, the isolated antigen-binding protein comprises an H-FR3 located between the HCDR2 and the HCDR3 and comprising the amino acid sequence represented by SEQ ID NO: 64.

[0017] In some embodiments, the H-FR3 comprises an amino acid sequence represented by any one of SEQ ID NO: 18 and SEQ ID NO: 29.

[0018] In some embodiments, the isolated antigen-binding protein comprises an H-FR4 having an N-terminus directly or indirectly linked to the C-terminus of the HCDR3 and comprising an amino acid sequence represented by SEQ ID NO: 65.

[0019] In some embodiments, the H-FR4 comprises an amino acid sequence represented by any one of SEQ ID NO: 20 and SEQ ID NO: 30.

[0020] In some embodiments, the isolated antigen-binding protein comprises H-FR1, H-FR2, H-FR3, and H-FR4, wherein the H-FR1 comprises an amino acid sequence represented by SEQ ID NO: 62, the H-FR2 comprises an amino acid sequence represented by SEQ ID NO: 63, the H-FR3 comprises an amino acid sequence represented by SEQ ID NO: 64, and the H-FR4 comprises an amino acid sequence represented by SEQ ID NO: 65.

[0021] In some embodiments, the H-FR1 comprises an amino acid sequence represented by any one of SEQ ID NO: 14 and SEQ ID NO: 27, the H-FR2 comprises an amino acid sequence represented by any one of SEQ ID NO: 16 and SEQ ID NO: 28, the H-FR3 comprises an amino acid sequence represented by any one of SEQ ID NO: 18 and SEQ ID NO: 29, and the H-FR4 comprises an amino acid sequence represented by any one of SEQ ID NO: 20 and SEQ ID NO: 30.

[0022] In some embodiments, in the isolated antigen-binding protein, the H-FR1, H-FR2, H-FR3, and H-FR4 comprise any one set of amino acid sequences selected from the following. a) H-FR1: SEQ ID NO: 14, H-FR2: SEQ ID NO: 16, H-FR3: SEQ ID NO: 18, and 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, and H-FR4: SEQ ID NO: 30

[0023] In some embodiments, the isolated antigen-binding protein comprises a heavy chain variable region VH comprising the amino acid sequence set forth in SEQ ID NO: 67.

[0024] In some embodiments, the isolated antigen-binding protein comprises a heavy chain variable region VH comprising the amino acid sequence set forth in either SEQ ID NO: 13 or SEQ ID NO: 26.

[0025] In some embodiments, the isolated antigen-binding protein comprises an LCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 11.

[0026] In some embodiments, the isolated antigen-binding protein comprises an LCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 9.

[0027] In some embodiments, the isolated antigen-binding protein comprises an LCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 7.

[0028] In some embodiments, the isolated antigen-binding protein comprises the LCDR1, LCDR2, and LCDR3 of a light chain variable region VL set forth in SEQ ID NO: 66.

[0029] In some embodiments, the isolated antigen-binding protein comprises the LCDR1, LCDR2, and LCDR3 of a light chain variable region VL set forth in either SEQ ID NO: 5 or 21.

[0030] In some embodiments, the isolated antigen-binding protein comprises a variable light chain region VL comprising the LCDR1, LCDR2, and LCDR3, wherein the LCDR3 comprises the amino acid 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.

[0031] In some embodiments, the isolated antigen-binding protein comprises an L-FR1 having a C-terminus directly or indirectly linked to the N-terminus of the LCDR1 and comprising the amino acid sequence shown in SEQ ID NO: 58.

[0032] In some embodiments, the L-FR1 comprises the amino acid sequence shown by either SEQ ID NO: 6 or SEQ ID NO: 22.

[0033] In some embodiments, the isolated antigen-binding protein comprises an L-FR2 located between the LCDR1 and the LCDR2 and comprising the amino acid sequence shown in SEQ ID NO: 59.

[0034] In some embodiments, the L-FR2 comprises the amino acid sequence shown by either SEQ ID NO: 8 or SEQ ID NO: 23.

[0035] In some embodiments, the isolated antigen-binding protein comprises an L-FR3 located between the LCDR2 and the LCDR3 and comprising the amino acid sequence shown in SEQ ID NO: 60.

[0036] In some embodiments, the L-FR3 comprises the amino acid sequence shown by either SEQ ID NO: 10 or SEQ ID NO: 24.

[0037] In some embodiments, the isolated antigen-binding protein comprises an L-FR4 having an N-terminus directly or indirectly linked to the C-terminus of the LCDR3 and comprising the amino acid sequence set forth in SEQ ID NO: 61.

[0038] In some embodiments, in the isolated antigen-binding protein, the L-FR4 comprises the amino acid sequence set forth in either SEQ ID NO: 12 or SEQ ID NO: 25.

[0039] In some embodiments, the isolated antigen-binding protein comprises L-FR1, L-FR2, L-FR3, and L-FR4, wherein the L-FR1 comprises the amino acid sequence set forth in SEQ ID NO: 58, the L-FR2 comprises the amino acid sequence set forth in SEQ ID NO: 59, the L-FR3 comprises the amino acid sequence set forth in SEQ ID NO: 60, and the L-FR4 comprises the amino acid sequence set forth in SEQ ID NO: 61.

[0040] In some embodiments, the L-FR1 comprises the amino acid sequence set forth in either SEQ ID NO: 6 or SEQ ID NO: 22, the L-FR2 comprises the amino acid sequence set forth in either SEQ ID NO: 8 or SEQ ID NO: 23, the L-FR3 comprises the amino acid sequence set forth in either SEQ ID NO: 10 or SEQ ID NO: 24, and the L-FR4 comprises the amino acid sequence set forth in either SEQ ID NO: 12 or SEQ ID NO: 25.

[0041] In some embodiments, in the isolated antigen-binding protein, the L-FR1, L-FR2, L-FR3, and L-FR4 comprise any one set of amino acid sequences selected from the following. a) 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 b) L-FR1: SEQ ID NO: 22, L-FR2: SEQ ID NO: 23, L-FR3: SEQ ID NO: 24, and L-FR4: SEQ ID NO: 25

[0042] In some embodiments, in the isolated antigen-binding protein, the VL comprises the amino acid sequence shown in SEQ ID NO: 66.

[0043] In some embodiments, the VL comprises the amino acid sequence shown in either SEQ ID NO: 5 or SEQ ID NO: 21.

[0044] In some embodiments, in the isolated antigen-binding protein, the VH and VL comprise any one set of amino acid sequences selected from the following. a) VH: SEQ ID NO: 13, and VL: SEQ ID NO: 5 b) VH: SEQ ID NO: 26, and VL: SEQ ID NO: 21

[0045] In some embodiments, the isolated antigen-binding protein comprises an HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 45.

[0046] In some embodiments, the isolated antigen-binding protein comprises an HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 43.

[0047] In some embodiments, the isolated antigen-binding protein comprises an HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 41.

[0048] In some embodiments, the isolated antigen-binding protein comprises HCDR1, HCDR2, and HCDR3 of the heavy chain variable region VH shown in SEQ ID NO: 77.

[0049] In some embodiments, the isolated antigen-binding protein comprises HCDR1, HCDR2, and HCDR3 of a heavy chain variable region VH represented by any of SEQ ID NO: 39 and SEQ ID NO: 52.

[0050] In some embodiments, the isolated antigen-binding protein comprises a heavy chain variable region VH comprising the HCDR1, HCDR2, and HCDR3, wherein the HCDR3 comprises the amino acid sequence represented by SEQ ID NO: 45, the HCDR2 comprises the amino acid sequence represented by SEQ ID NO: 43, and the HCDR1 comprises the amino acid sequence represented by SEQ ID NO: 41.

[0051] In some embodiments, the isolated antigen-binding protein comprises an H-FR1 having a C-terminus directly or indirectly linked to the N-terminus of the HCDR1 and comprising the amino acid sequence represented by SEQ ID NO: 72.

[0052] In some embodiments, in the isolated antigen-binding protein, the H-FR1 comprises the amino acid sequence represented by any of SEQ ID NO: 40 and SEQ ID NO: 53.

[0053] In some embodiments, the isolated antigen-binding protein comprises an H-FR2 located between the HCDR1 and the HCDR2 and comprising the amino acid sequence represented by SEQ ID NO: 73.

[0054] In some embodiments, in the isolated antigen-binding protein, the H-FR2 comprises the amino acid sequence represented by any of SEQ ID NO: 42 and SEQ ID NO: 54.

[0055] In some embodiments, the isolated antigen-binding protein comprises an H-FR3 located between the HCDR2 and the HCDR3 and comprising the amino acid sequence represented by SEQ ID NO: 74.

[0056] In some embodiments, in the isolated antigen-binding protein, the H-FR3 comprises an amino acid sequence represented by any one of SEQ ID NO: 44 and SEQ ID NO: 55.

[0057] In some embodiments, the isolated antigen-binding protein comprises an H-FR4 having an N-terminus directly or indirectly linked to the C-terminus of the HCDR3 and comprising an amino acid sequence represented by SEQ ID NO: 75.

[0058] In some embodiments, the isolated antigen-binding protein comprises an H-FR4 having an N-terminus directly or indirectly linked to the C-terminus of the HCDR3 and comprising an amino acid sequence represented by any one of SEQ ID NO: 46 and SEQ ID NO: 30.

[0059] In some embodiments, the isolated antigen-binding protein comprises H-FR1, H-FR2, H-FR3, and H-FR4, wherein the H-FR1 comprises an amino acid sequence represented by SEQ ID NO: 72, the H-FR2 comprises an amino acid sequence represented by SEQ ID NO: 73, the H-FR3 comprises an amino acid sequence represented by SEQ ID NO: 74, and the H-FR4 comprises an amino acid sequence represented by SEQ ID NO: 75.

[0060] In some embodiments, in the isolated antigen-binding protein, the H-FR1 comprises an amino acid sequence represented by any one of SEQ ID NO: 40 and SEQ ID NO: 53, the H-FR2 comprises an amino acid sequence represented by any one of SEQ ID NO: 42 and SEQ ID NO: 54, the H-FR3 comprises an amino acid sequence represented by any one of SEQ ID NO: 44 and SEQ ID NO: 55, and the H-FR4 comprises an amino acid sequence represented by any one of SEQ ID NO: 46 and SEQ ID NO: 30.

[0061] In some embodiments, in the isolated antigen-binding protein, the H-FR1, H-FR2, H-FR3, and H-FR4 comprise any one set of amino acid sequences selected from the following. a) H-FR1: SEQ ID NO: 40, H-FR2: SEQ ID NO: 42, H-FR3: SEQ ID NO: 44, and 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, and H-FR4: SEQ ID NO: 30

[0062] In some embodiments, the isolated antigen-binding protein comprises a heavy-chain variable region VH comprising the amino acid sequence shown by SEQ ID NO: 77.

[0063] In some embodiments, in the isolated antigen-binding protein, the VH comprises the amino acid sequence shown by any one of SEQ ID NO: 39 and SEQ ID NO: 52.

[0064] In some embodiments, the isolated antigen-binding protein comprises an LCDR3 comprising the amino acid sequence shown by SEQ ID NO: 37.

[0065] In some embodiments, the isolated antigen-binding protein comprises an LCDR2 comprising the amino acid sequence shown by SEQ ID NO: 35.

[0066] In some embodiments, the isolated antigen-binding protein comprises an LCDR1 comprising the amino acid sequence shown by SEQ ID NO: 33.

[0067] In some embodiments, the isolated antigen-binding protein comprises the LCDR1, LCDR2, and LCDR3 of a light-chain variable region VL shown by SEQ ID NO: 76.

[0068] In some embodiments, the isolated antigen-binding protein comprises LCDR1, LCDR2, and LCDR3 of the variable light chain region VL represented by any of SEQ ID NO: 31 and SEQ ID NO: 47.

[0069] In some embodiments, the isolated antigen-binding protein comprises a variable light chain region VL comprising the LCDR1, LCDR2, and LCDR3, wherein the LCDR3 comprises the amino acid sequence represented by SEQ ID NO: 37, the LCDR2 comprises the amino acid sequence represented by SEQ ID NO: 35, and the LCDR1 comprises the amino acid sequence represented by SEQ ID NO: 33.

[0070] In some embodiments, the isolated antigen-binding protein comprises L-FR1 having a C-terminus directly or indirectly linked to the N-terminus of the LCDR1 and comprising the amino acid sequence represented by SEQ ID NO: 68.

[0071] In some embodiments, in the isolated antigen-binding protein, the L-FR1 comprises the amino acid sequence represented by any of SEQ ID NO: 32 and SEQ ID NO: 48.

[0072] In some embodiments, the isolated antigen-binding protein comprises L-FR2 located between the LCDR1 and the LCDR2 and comprising the amino acid sequence represented by SEQ ID NO: 69.

[0073] In some embodiments, in the isolated antigen-binding protein, the L-FR2 comprises the amino acid sequence represented by any of SEQ ID NO: 34 and SEQ ID NO: 49.

[0074] In some embodiments, the isolated antigen-binding protein comprises L-FR3 located between the LCDR2 and the LCDR3 and comprising the amino acid sequence represented by SEQ ID NO: 70.

[0075] In some embodiments, in the isolated antigen-binding protein, the L-FR3 comprises an amino acid sequence represented by any one of SEQ ID NO: 36 and SEQ ID NO: 50.

[0076] In some embodiments, the isolated antigen-binding protein comprises an L-FR4 having an N-terminus directly or indirectly linked to the C-terminus of the LCDR3 and comprising an amino acid sequence represented by SEQ ID NO: 71.

[0077] In some embodiments, in the isolated antigen-binding protein, the L-FR4 comprises an amino acid sequence represented by any one of SEQ ID NO: 38 and SEQ ID NO: 51.

[0078] In some embodiments, the isolated antigen-binding protein comprises L-FR1, L-FR2, L-FR3, and L-FR4, wherein the L-FR1 comprises an amino acid sequence represented by SEQ ID NO: 68, the L-FR2 comprises an amino acid sequence represented by SEQ ID NO: 69, the L-FR3 comprises an amino acid sequence represented by SEQ ID NO: 70, and the L-FR4 comprises an amino acid sequence represented by SEQ ID NO: 71.

[0079] In some embodiments, in the isolated antigen-binding protein, the L-FR1 comprises an amino acid sequence represented by any one of SEQ ID NO: 32 and SEQ ID NO: 48, the L-FR2 comprises an amino acid sequence represented by any one of SEQ ID NO: 34 and SEQ ID NO: 49, the L-FR3 comprises an amino acid sequence represented by any one of SEQ ID NO: 36 and SEQ ID NO: 50, and the L-FR4 comprises an amino acid sequence represented by any one of SEQ ID NO: 38 and SEQ ID NO: 51.

[0080] In some embodiments, in the isolated antigen-binding protein, the L-FR1, L-FR2, L-FR3, and L-FR4 comprise any one set of amino acid sequences selected from the following. a) L-FR1: SEQ ID NO: 32, L-FR2: SEQ ID NO: 34, L-FR3: SEQ ID NO: 36, and 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, and L-FR4: SEQ ID NO: 51

[0081] In some embodiments, the isolated antigen-binding protein comprises a VL comprising the amino acid sequence represented by SEQ ID NO: 76.

[0082] In some embodiments, in the isolated antigen-binding protein, the VL comprises the amino acid sequence represented by any one of SEQ ID NO: 31 and 47.

[0083] In some embodiments, the isolated antigen-binding protein comprises a VH and a VL, and the VH and VL comprise any one set of amino acid sequences selected from the following. a) VH: SEQ ID NO: 39, and VL: SEQ ID NO: 31 b) VH: SEQ ID NO: 52, and VL: SEQ ID NO: 47

[0084] In some embodiments, the isolated antigen-binding protein comprises a heavy chain constant region comprising a constant region derived from IgG or a constant region derived from IgY.

[0085] In some embodiments, in the isolated antibody-binding protein, the heavy chain constant region comprises a constant region derived from IgG.

[0086] In some embodiments, in the isolated antigen-binding protein, the heavy chain constant region comprises a constant region derived from IgG1, IgG2, IgG3, or IgG4.

[0087] In some embodiments, in the isolated antigen-binding protein, the heavy chain constant region comprises the amino acid sequence shown in SEQ ID NO: 56.

[0088] In some embodiments, the isolated antigen-binding protein comprises a light chain constant region comprising a constant region derived from Igκ or a constant region derived from Igλ.

[0089] In some embodiments, in the isolated antigen-binding protein, the light chain constant region comprises a constant region derived from human Igκ.

[0090] In some embodiments, in the isolated antigen-binding protein, the light chain constant region comprises the amino acid sequence shown in SEQ ID NO: 57.

[0091] In some embodiments, the isolated antigen-binding protein comprises an antibody or an antigen-binding fragment thereof.

[0092] In some embodiments, in the isolated antigen-binding protein, the antigen-binding fragment is selected from the group consisting of Fab, Fab’, F(ab)2, Fv fragment, F(ab’)2, scFv, di-scFv, VHH, and / or dAb.

[0093] In some embodiments, in the isolated antigen-binding protein, the antibody is selected from the group consisting of monoclonal antibodies, single-chain antibodies, chimeric antibodies, humanized antibodies, and fully human antibodies.

[0094] In another aspect, the present application provides one or more polypeptides comprising the isolated antigen-binding protein.

[0095] In another aspect, the present application provides one or more immunoconjugates comprising the isolated antigen-binding protein or the polypeptide.

[0096] In another aspect, the present application provides one or more isolated nucleic acid molecules encoding the isolated antigen-binding protein or the polypeptide.

[0097] In another aspect, the present application provides one or more vectors comprising the isolated nucleic acid molecule.

[0098] In another aspect, the present application provides one or more cells comprising the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, and / or the vector.

[0099] In another aspect, the present application provides a method for producing the isolated antigen-binding protein or the polypeptide, comprising culturing the cells under conditions for expressing the isolated antigen-binding protein or the polypeptide.

[0100] In another aspect, the present application provides one or more pharmaceutical compositions comprising the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, the cell, and / or a pharmaceutically acceptable adjuvant and / or excipient.

[0101] In another aspect, the present application provides a method for detecting or measuring MASP-2, comprising using the isolated antigen-binding protein or the polypeptide.

[0102] In another aspect, the present application provides a detection kit for MASP-2, comprising the isolated antigen-binding protein or the polypeptide.

[0103] In another aspect, the present application provides the use of the isolated antigen-binding protein or the polypeptide in the manufacture of a kit for detecting the presence and / or content of MASP-2.

[0104] In another aspect, the present application provides the use of the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, the cell and / or the pharmaceutical composition in the manufacture of a drug for preventing and / or treating a disease or disorder.

[0105] In another aspect, the present application provides the use of the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, the cell and / or the pharmaceutical composition in the prevention, remission and / or treatment of a disease or disorder.

[0106] In another aspect, the present application provides a method for preventing and / or treating a disease or disorder, comprising administering an effective amount of the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, and / or the cell to a subject in need thereof.

[0107] Those skilled in the art can easily grasp other aspects and advantages of the present application from the following detailed description. In the following detailed description, only exemplary embodiments of the present application are shown and described. As those skilled in the art will recognize, the content of the present application allows those skilled in the art to modify the disclosed specific embodiments without departing from the spirit and scope of the invention according to the present application. Correspondingly, the description of the drawings and the specification of the present application are merely exemplary and not limiting.

Brief Description of the Drawings

[0108] Specific features of the invention according to the present application are shown in the appended claims. The features and advantages of the invention according to the present application can be better understood by referring to the exemplary embodiments described in detail below and the appended drawings. The drawings are briefly described as follows

[0109]

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Modes for Carrying Out the Invention

[0110] Hereinafter, embodiments of the present application will be described by specific specific examples. However, those skilled in the art can easily understand other advantages and effects of the present application from the content disclosed in this specification. Term Definitions

[0111] As used herein, the term "isolated" generally means obtained by artificial means from its natural state. If an "isolated" substance or component appears in nature, the natural environment in which it is placed may have changed, the substance may have been isolated from its natural environment, or both situations may have occurred. For example, a certain unisolated polynucleotide or polypeptide naturally exists in the body of a living animal, and the same high-purity polynucleotide or polypeptide isolated from its natural state is referred to as an isolated one. The term "isolated" does not exclude the presence of artificial or synthetic substances or other impurities that do not affect the activity of the substance.

[0112] In the present application, the term "antigen-binding protein" generally refers to a polypeptide molecule that can specifically recognize and / or neutralize a specific antigen. For example, in the present application, the term "antigen-binding protein" may include an "antibody" or an "antigen-binding fragment". For example, the antibody may include an immunoglobulin composed of at least two heavy (H) chains and two light (L) chains linked to each other via disulfide bonds, and may include any molecule containing its antigen-binding portion. The term "antibody" includes, but is not limited to, monoclonal antibodies, antibody fragments or antibody derivatives such as mouse-derived antibodies, human antibodies (fully human antibodies), humanized antibodies, chimeric antibodies, single-chain antibodies (e.g., scFv), and antibody fragments that bind to an antigen (e.g., Fab, Fab', VHH, and (Fab)2 fragments). The term "antibody" may also include all recombinant forms of antibodies, such as antibodies expressed in prokaryotic cells, non-glycosylated antibodies, and any antibody fragments and their derivatives that bind to the antigens described herein. Each heavy chain may be composed of a heavy chain variable region (VH) and a heavy chain constant region. Each light chain may be composed of a light chain variable region (VL) and a light chain constant region. The VH and VL regions may be further distinguished as hypervariable regions called complementarity-determining regions (CDRs), which are interspersed among more conserved regions called framework regions (FRs). Each VH and VL may be composed of three CDR regions and four FR regions, and may be arranged in the order of FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4 from the amino terminus to the carboxyl terminus. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen (e.g., human MASP-2). The constant region of the antibody can mediate the binding of the immunoglobulin to host tissues or factors including multiple types of cells of the immune system (e.g., effector cells) and the first component of the classical complement system (Clq). The exact boundaries of the CDRs are defined differently according to different systems.The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991))) provides not only a clear residue numbering system applicable to any variable region of an antigen-binding fragment, but also the exact residue boundaries that define the CDRs. These CDRs can be referred to as Kabat CDRs. Chothia and colleagues (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987), and Chothia et al., Nature 342:877-883 (1989)) discovered that, despite great diversity at the amino acid sequence level, some subsections within the Kabat CDRs adopt nearly the same peptide backbone conformation. These subsections are named L1, L2, and L3, or H1, H2, and H3, where "L" and "H" refer to the light-chain and heavy-chain regions, respectively. These regions can be referred to as Chothia CDRs, which have boundaries overlapping with the Kabat CDRs. Other boundaries that overlap with the Kabat CDRs and define the CDRs are described by Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol 262(5):732-45 (1996)). Also, definitions of other CDR boundaries may not strictly follow one of the above systems but still overlap with the Kabat CDRs. However, they can be shortened or extended according to predictions or experimental findings that a particular residue or group of residues, or the CDR as a whole, does not have a significant impact on antigen binding. In the present application, the CDRs may be defined using the Chothia numbering system.

[0113] In the present application, the term "antigen-binding fragment" generally means one or more fragments of an antibody that exhibit the function of specifically binding to an antigen. The antigen-binding function of an antibody can be achieved by the full-length fragment of the antibody. The antigen-binding function of an antibody can also be achieved by a heavy chain containing a fragment of Fv, ScFv, dsFv, Fab, Fab' or F(ab')2, or a light chain containing a fragment of Fv, scFv, dsFv, Fab, Fab' or F(ab')2. (1) Generally, a Fab fragment which is a monovalent fragment consisting of VL, VH, CL and CH domains, (2) an F(ab')2 fragment which is a divalent fragment containing two Fab fragments linked via a disulfide bond in the hinge region, (3) an Fd fragment consisting of VH and CH domains, (4) an Fv fragment consisting of VL and VH domains of a single arm of an antibody, (5) a dAb fragment consisting of a VH domain (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 that can be optionally linked via a linker. For example, it may also include a monovalent single-chain molecule Fv (scFv) formed from 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). For example, an antibody VHH that lacks the light chain of the antibody and has only the variable region of the heavy chain (see, for example, Kang Xiaosen et al., Biotechnology Journal 2018, 34(12): 1974-1984). The "antigen-binding site" may further include an immunoglobulin fusion protein containing the following binding domains. (1) A binding domain polypeptide fused to an immunoglobulin hinge region polypeptide, (2) 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.

[0114] In the present application, the term "monoclonal antibody" generally means a collection of substantially identical antibodies, i.e., the individual antibodies contained in this collection are identical except for naturally occurring mutations that may be present in trace amounts. Monoclonal antibodies are highly specific and directly target a single antigenic site. For example, the monoclonal antibody can be prepared by hybridoma technology or produced in bacteria, eukaryotes or plant cells using recombinant DNA methods. Monoclonal antibodies may also be obtained from phage antibody libraries using, for example, the techniques described in Clackson et al., Nature, 352:624-628 (1991) and Marks et al., Mol. Biol., 222:581-597 (1991).

[0115] In the present application, the term "chimeric antibody" generally means an antibody in which a part of the amino acid sequence of each heavy or light chain is homologous to the corresponding amino acid sequence of an antibody derived from a specific species or belongs to a specific class, and the remaining part of that chain is homologous to the corresponding sequence of another species. For example, the variable regions of both the light and heavy chains are derived from the variable regions of antibodies of the same animal species (e.g., mouse, rat, etc.), and the constant part is homologous to an antibody sequence derived from another species (e.g., human). For example, to obtain a chimeric antibody, variable regions can be produced using non-human-derived B cells or hybridoma cells, and the constant regions combined with them are of human origin. The variable regions have the advantage of being easily prepared, and their specificity is not affected by the origin of the constant regions combined with them. On the other hand, since the constant regions of chimeric antibodies may be of human origin, chimerics are less likely to induce an immune response upon injection than antibodies with non-human-derived constant regions.

[0116] As used herein, the term "humanized antibody" generally refers to a chimeric antibody that has a reduced number of sequences derived from non-human immunoglobulins, thereby reducing the immunogenicity when a heterologous antibody is introduced into a human, while maintaining the full antigen-binding affinity and specificity of the antibody. For example, techniques such as CDR grafting (Jones et al., Nature 321:522 (1986)) and its variants, "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), "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. Cellular adhesion: molecular definition to therapeutic potential. New York: Plenum Press, 1994: 291-312), surface reconstruction (U.S. Patent No. US5639641), etc. are used to humanize the non-human-derived binding domain. If other regions such as the hinge region and the constant region domain are also of non-human origin, these regions can also be humanized.

[0117] As used herein, the term "mouse-derived antibody" generally means an antibody in which the variable region framework and CDR regions are derived from mouse germline immunoglobulin sequences. Also, when the antibody includes a constant region, it is also obtained from mouse germline immunoglobulin sequences. The mouse-derived antibodies of the present application can contain amino acid residues not encoded by mouse germline immunoglobulin sequences, and can contain, for example, mutations introduced by random mutagenesis or point mutagenesis in vitro, or by somatic mutations in vivo.

[0118] As used herein, the term "germline sequence" generally means the sequence of an un-rearranged immunoglobulin DNA sequence.

[0119] As used herein, the term "between" generally means that the C-terminus of a certain amino acid fragment is directly or indirectly linked to the N-terminus of the first amino acid fragment, and its N-terminus is directly or indirectly linked to the C-terminus of the second amino acid fragment. In the light chain, for example, the N-terminus of the L-FR2 is directly or indirectly linked to the C-terminus of the LCDR1, and the C-terminus of the L-FR2 is directly or indirectly linked to the N-terminus of the LCDR2. Further, for example, the N-terminus of the L-FR3 is directly or indirectly linked to the C-terminus of the LCDR2, and the C-terminus of the L-FR3 is directly or indirectly linked to the N-terminus of the LCDR3. In the heavy chain, for example, the N-terminus of the H-FR2 is directly or indirectly linked to the C-terminus of the HCDR1, and the C-terminus of the H-FR2 is directly or indirectly linked to the N-terminus of the HCDR2. Further, for example, the N-terminus of the H-FR3 is directly or indirectly linked to the C-terminus of the HCDR2, and the C-terminus of the H-FR3 is directly or indirectly linked to the N-terminus of the HCDR3.

[0120] In the present application, the terms "MASP-2 protein", "MASP-2", or "MASP-2 antigen" can be used interchangeably and include any functionally active fragment, variant, and homolog of MASP-2, which is either naturally expressed by cells or expressed on cells transfected with the MASP-2 gene. In the present application, MASP-2 may be human MASP-2 with the accession number O00187 in UniProt / Swiss-Prot. For example, MASP-2 can be a functionally active fragment of human MASP-2. In the present application, MASP-2 can be cynomolgus monkey MASP-2 or a functionally active fragment thereof. For example, the "functionally active fragment" may include a fragment that retains the endogenous function of at least one naturally occurring protein (e.g., binds to the antigen-binding protein according to the present application). For example, the "functionally active fragment" may include a domain that binds to the antigen-binding protein of the present application. In the present application, MASP-2 is an important regulatory factor in the lectin pathway of the complement system. Mannan-binding lectin (MBL) and ficolin (FCN) in plasma directly recognize sugar structures with terminal sugar groups such as mannose, N-acetylmannose, N-acetylglucosamine, and fucose on the surfaces of various pathogenic microorganisms. The MBL-MASP complex binds to the sugar structure on the pathogen surface and independently activates MASP-1 and MASP-2. Activated MASP-2 exerts its SP activity, cleaves C4, and the generated C4b fragment covalently binds to the pathogen surface. Through interaction with C2, the latter is also cleaved by MASP-2 to form the C3 convertase C4b2a, subsequently activating the classical complement pathway. Activated MASP1 can directly cleave C3 to produce C3b, and under the action of protein factor D and protein factor P, form the C3 convertase C3bBb or C3bBbP, and produce the C5 convertase C3bBb3b to activate the alternative complement pathway.

[0121] In the present application, the term "IgA nephropathy" generally means primary glomerulopathy in the glomerular mesangial region. For example, the "IgA nephropathy" may include primary glomerulopathy mainly characterized by deposition of IgA or IgA with or without deposition of other immunoglobulins in the glomerular mesangial region. For example, the lesion types of the "IgA nephropathy" may include focal segmental lesions, endocapillary proliferative lesions, mesangial proliferative lesions, crescentic lesions, sclerotic lesions, and the like. Clinical symptoms include recurrent gross hematuria and microscopic hematuria accompanied by varying degrees of proteinuria, and some patients may develop severe hypertension or renal insufficiency.

[0122] In addition to the specific proteins and nucleotides described herein, the present application can include functionally active fragments, derivatives, analogs, homologs, and fragments thereof.

[0123] The term "functionally active fragment" means a polypeptide having an amino acid sequence substantially identical to a naturally occurring sequence or encoded by a substantially identical nucleotide sequence and capable of having one or more activities of the naturally occurring sequence. In the context of the present application, a functionally active fragment of any given sequence means a sequence in which the specific sequence of its residues (whether amino acid residues or nucleotide residues) is modified such that the polypeptide or polynucleotide substantially retains at least one endogenous function. By addition, deletion, replacement, modification, substitution, and / or mutation of at least one amino acid residue and / or nucleotide residue present in a naturally occurring protein and / or polynucleotide, a sequence encoding a functionally active fragment can be obtained, as long as the original functional activity can be maintained.

[0124] In the present application, the term "derivative" generally means a polypeptide or polynucleotide of the present application that includes any replacement, mutation, modification, substitution, deletion, and / or addition of one (or more) amino acid residues from / to a sequence, provided that the resulting polypeptide or polynucleotide can substantially retain at least one endogenous function thereof.

[0125] In the present application, the term "analog" generally includes, for a polypeptide or polynucleotide, any analog of the polypeptide or polynucleotide, i.e., a chemical compound having at least one endogenous function of the polypeptide or polynucleotide that has been analogized by the analog.

[0126] Generally, amino acid substitutions, for example, at least one (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 20 or more) amino acid substitutions can be made as long as the modified sequence substantially maintains the desired activity or ability. Amino acid substitutions may include the use of non-naturally occurring analogs.

[0127] As used herein, the term "homolog" generally refers to an amino acid sequence or nucleotide sequence having a certain homology with a naturally occurring sequence. The term "homology" may be equivalent to sequence "identity". Homologous sequences may include amino acid sequences that are at least 80%, at least 85%, at least 90%, at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8% or at least 99.9% identical to a target sequence. Generally, homologs include the same active site as the target amino acid sequence, etc. Homology can be considered based on similarity (i.e., amino acid residues with similar chemical properties and functions), and homology can also be exhibited in sequence identity. As used herein, a sequence having a predetermined percentage identity with any of the SEQ ID NOs of the recited amino acid sequences or nucleotide sequences refers to a sequence having that percentage identity over the entire length of the relevant SEQ ID NO. To determine sequence identity, sequence alignment can be performed, and sequence alignment can be performed in various ways known to those skilled in the art using, for example, BLAST, BLAST-2, ALIGN, NEEDLE or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for alignment, including the algorithms necessary to achieve optimal alignment in the compared full-length sequences.

[0128] The proteins or polypeptides used in the present application may have deletions, insertions or substitutions of amino acid residues that result in silent changes and yield functionally equivalent proteins. Intentional amino acid substitutions may be made according to the similarity of the polarity, charge, solubility, hydrophobicity, hydrophilicity and / or amphoteric properties of the residues, as long as the endogenous function is retained. For example, negatively charged amino acids include aspartic acid and glutamic acid, and positively charged amino acids include lysine and arginine. In addition, amino acids having similar hydrophilicity values and no polar head groups include asparagine, glutamine, serine, threonine and tyrosine.

[0129] As used herein, the term "immunoconjugate" generally refers to a conjugate formed by the complexation (e.g., covalent bonding by a linking molecule) of said other therapeutic agent with said isolated antigen-binding protein, which conjugate can deliver said other therapeutic agent to target cells by specific binding of said isolated antigen-binding protein to an antigen on the target cells. Also, said antigen can be secreted from said target cells and can be located in the extracellular space outside said target cells.

[0130] As used herein, the term "subject" generally refers to a human or non-human animal, including but not limited to cats, dogs, horses, pigs, cows, sheep, rabbits, mice, rats, or monkeys.

[0131] As used herein, the term "nucleic acid molecule" generally refers to an isolated form of nucleotides, deoxyribonucleotides, or ribonucleotides or their analogs of any length, isolated from its natural environment or artificially synthesized.

[0132] As used herein, the term "vector" generally refers to a nucleic acid molecule capable of transporting another nucleic acid linked thereto. Said vector can introduce the inserted nucleic acid molecule into cells and / or between cells. Said vector may include a vector mainly for inserting DNA or RNA into cells, a vector mainly for replicating DNA or RNA, and a vector mainly for expression for transcription and / or translation of DNA or RNA. Said vector may be a polynucleotide capable of being transcribed and translated into a polypeptide when an appropriate cell is introduced. Generally, a vector can produce a desired expression product by culturing an appropriate cell containing said vector. As used herein, said vector may include a lentiviral vector.

[0133] In the present application, the term "cell" generally can include, or already includes, a plasmid or vector containing a nucleic acid molecule according to the present application, or an individual cell, cell line, or cell culture capable of expressing a polypeptide or antigen-binding protein according to the present application. The cell may include progeny of a single cell. Due to natural, unexpected, or intentional mutations, the progeny cells may not necessarily be completely identical morphologically and genomically to the parental cells, but it is sufficient if they can express the polypeptide or antigen-binding protein according to the present application. The cell can be obtained by transfecting cells in vitro using a vector according to the present application. The cell may be a prokaryotic cell (e.g., Escherichia coli), or a eukaryotic cell (e.g., a yeast cell, e.g., a COS cell, a Chinese hamster ovary (CHO) cell, a HeLa cell, a HEK293 cell, a COS-1 cell, an NS0 cell, or a myeloma cell). In some embodiments, the cell may be an immune cell. For example, the immune cell can be selected from T cells, B cells, natural killer cells (NK cells), macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, leukocytes, and / or peripheral blood monocytes.

[0134] In the present application, the term "treatment" generally refers to (i) preventing the onset of a disease, disorder, or symptom in a patient who is at risk of developing it but has not been diagnosed with it, (ii) suppressing the disease, disorder, or symptom, i.e., suppressing its progression, and (iii) alleviating the disease, disorder, or symptom, i.e., eliminating the disease, disorder, and / or symptom and / or symptoms associated with the disease, symptom, and / or symptoms.

[0135] In the present application, the terms "polypeptide", "peptide", "protein" and "protein" are used interchangeably and generally mean a polymer having amino acids of any length. This polymer may be linear or branched, may contain modified amino acids, and may be interrupted by non-amino acids. These terms also include modified amino acid polymers. These modifications may include disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other operation (such as binding to a labeling component). The term "amino acid" includes natural and / or non-natural or synthetic amino acids, including glycine, D and L enantiomers, as well as amino acid analogs and peptide analogs.

[0136] In the present application, the terms "polynucleotide", "nucleotide", "nucleotide sequence", "nucleic acid", and "oligonucleotide" are used interchangeably and generally mean a polymeric form of nucleotides of any length, such as deoxyribonucleotides, ribonucleotides, or their analogs. The polynucleotide may have any three-dimensional structure and may perform any known or unknown function. Non-limiting examples of polynucleotides include the coding or non-coding regions of a gene or gene fragment, multiple loci (one locus) defined by junction analysis, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, short interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA), ribozyme, cDNA, recombinant polynucleotide, branched polynucleotide, plasmid, vector, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probe, and primer. The polynucleotide may contain one or more modified nucleotides such as methylated nucleotides and nucleotide analogs. Modification of the nucleotide structure, if present, may be performed before or after assembly of the polymer. The nucleotide sequence may be interrupted by non-nucleotide components. The polynucleotide may be further modified after polymerization, such as by conjugation with a labeled component.

[0137] In the present application, "K" D " (similarly, "K" D " or "K" D ") generally refers to the "affinity constant" or "equilibrium dissociation constant" and means the value obtained by dividing the equilibrium value or dissociation rate constant (kd) by the association rate constant (ka) in titration measurements. The association rate constant (ka), dissociation rate constant (kd), and equilibrium dissociation constant (K" D( ) represents the binding affinity of a binding protein (e.g., the isolated antigen-binding protein according to the present application) for an antigen (e.g., MASP-2 protein). Methods for determining the binding and dissociation rate constants are well known to those skilled in the art. By using fluorescence-based techniques, high sensitivity is provided and it is possible to examine the sample when it is in equilibrium in a physiological buffer. The said K D value can be measured, for example, by Biacore (biomolecular interaction analysis) (e.g., with an apparatus available from BIAcore International AB, a GE Healthcare company, Uppsala, Sweden), and can also be detected using other experimental routes and instruments such as Octet detection. Also, the said K D value can also be measured using KinExA (Kinetic Exclusion Assay) obtained from Sapidyne Instruments (Boise, Idaho), and can also be measured using a surface plasmon resonance apparatus (SPR). The said K D value can also be measured, for example, by an amine coupling kit.

[0138] In the present application, the term "and / or" is understood to mean either option or both options.

[0139] In the present application, the term "comprising" generally means including other elements without excluding them, but including the specifically designated features. In some cases, "comprising" may mean including only the designated components. For example, it has the meaning of "is", or also has the meaning of "consisting of".

[0140] In the present application, the term "about" generally means a variation within the range of 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%, or 10% or more or less of the designated value, that is, within the range of 0.5% to 10% or more or less of the designated value.

[0141] In the present application, the term "comprising" generally refers to the meaning of "including", "containing", "having", or "covering". In some cases, it also has the meaning of "being" or "consisting of".

[0142] Isolated antigen-binding protein according to the present application In one aspect, the present application provides an isolated antigen-binding protein that can specifically bind to human MASP-2 protein with a K D value of about 2E-09 M or less (for example, the K D is about 2E-09 M or less, about 1.5E-09 M or less, about 1E-09 M or less, about 9E-10 M or less, about 8E-10 M or less, about 7E-10 M or less, about 6E-10 M or less, about 5E-10 M or less, about 2E-10 M or less, about 1E-10 M or less, about 5E-11 M or less, about 1E-11 M or less, or 5E-12 M or less).

[0143] In one aspect, the present application provides an isolated antigen-binding protein that can specifically bind to cynomolgus monkey MASP-2 protein with a K D value of about 2E-09 M or less (for example, the K D is about 2E-09 M or less, about 1.5E-09 M or less, about 1E-09 M or less, about 9E-10 M or less, about 8E-10 M or less, about 7E-10 M or less, about 6E-10 M or less, about 5E-10 M or less, about 2E-10 M or less, about 1E-10 M or less, about 5E-11 M or less, about 1E-11 M or less, or 5E-12 M or less).

[0144] In one aspect, the present application provides an isolated antigen-binding protein that may comprise at least one CDR within the variable heavy region VH of an antibody, wherein the VH may comprise the amino acid sequence represented by SEQ ID NO: 67 or SEQ ID NO: 77.

[0145] For example, the VH may include an amino acid sequence represented by any of SEQ ID NO: 13, 26, 39, and 52. In the present application, the HCDR of the isolated antigen-binding protein may be divided in any form, and as long as the VH is homologous to the amino acid sequence represented by any of SEQ ID NO: 13, 26, 39, and 52, the HCDR divided in any form is within the scope of the patent of the present application.

[0146] The CDR of an antibody, also called a complementarity-determining region, is part of the variable region. The amino acid residues in this region may contact an antigen or an antigen epitope. The antibody CDR may be determined by various numbering systems such as CCG, Kabat, Chothia, IMGT, AbM, and both Kabat / Chothia. These numbering systems are known in the art, and specifically, for example, refer to http: / / www.bioinf.org.uk / abs / index.html#kabatnum. A person skilled in the art may determine the CDR region with various numbering systems according to the sequence and structure of the antibody. Depending on the numbering system used, there may be differences in the CDR region. In the present application, the CDR includes a CDR sequence divided by any CDR splitting method and variants thereof, and the variants include those obtained by substituting, deleting, and / or adding one or more amino acids to the amino acid sequence of the CDR, for example, 1 to 30, 1 to 20, or 1 to 10, and further, for example, those obtained by substituting, deleting, and / or inserting 1, 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, and also includes homologs thereof, and the homologs may be amino acid sequences having at least about 85% (for example, at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) sequence identity to the amino acid sequence of the CDR. In some embodiments, the isolated antigen-binding protein according to the present application is defined by the Chothia numbering system.

[0147] In the present application, the antigen-binding protein may include a heavy-chain variable region VH, and the VH may include at least one, two, or three of HCDR1, HCDR2, and HCDR3.

[0148] In the present application, the HCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 19. For example, the HCDR3 sequence of the antigen-binding protein may be defined by the Chothia numbering system.

[0149] In the present application, the HCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 17. For example, the HCDR2 sequence of the antigen-binding protein may be defined by the Chothia numbering system.

[0150] In the present application, the HCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 15. For example, the HCDR1 sequence of the antigen-binding protein may be defined by the Chothia numbering system.

[0151] For example, the HCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 15, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 17, and the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 19. For example, the antigen-binding protein may include antibody 50A6, JYB1931A63, or an antigen-binding fragment having the same HCDR3 (for example, the same HCDR1-3) as these.

[0152] For example, the VH of the antigen-binding protein may include framework regions H-FR1, H-FR2, H-FR3, and H-FR4.

[0153] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 62. For example, the H-FR1 of the antigen-binding protein has amino acid substitutions (such as conservative amino acid substitutions, etc.) at one or more amino acids selected from the group consisting of X 16 and X 19 compared with the sequence represented by SEQ ID NO: 62.

[0154] EVQLVESGGGLVQPGX 16 SLX 19 LSCAAS (SEQ ID NO: 62) (where X 16 may be G or R, and X 19 may be R or S)

[0155] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 14 and SEQ ID NO: 27.

[0156] In the present application, the H-FR2 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 63. For example, the H-FR2 of the antigen-binding protein has amino acid substitutions (such as conservative amino acid substitutions, etc.) at one or more amino acids selected from the group consisting of X 8 and X 10 compared with the sequence represented by SEQ ID NO: 63.

[0157] NMAWVRQX 8 PX 10 KGLEWVATI (SEQ ID NO: 63) (where X 8 may be A or T, and X 10 may be G or K)

[0158] In the present application, the H-FR2 of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 16 and SEQ ID NO: 28.

[0159] In the present application, the H-FR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 64. For example, compared with the sequence represented by SEQ ID NO: 64, the H-FR3 of the antigen-binding protein has an amino acid substitution (e.g., conservative amino acid substitution, etc.) at one or more amino acids selected from the group consisting of X 16 , X 20 , X 21 , X 27 , X 31 and X 36 .

[0160] TYYRDSVKGRFTISRX 16 NAKX 20 X 21 LYLQMX 27 SLRX 31 EDTAX 36 YYCST (SEQ ID NO: 64) (where X 16 may be D or E, X 20 may be N or S, X 21 may be S or T, X 27 may be D or N, X 31 may be A or S, X 36 may be T or V)

[0161] In the present application, the H-FR3 of the antigen-binding protein may include the amino acid sequence represented by any one of SEQ ID NO: 18 and SEQ ID NO: 29.

[0162] In the present application, the H-FR4 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 65. For example, compared with the sequence represented by SEQ ID NO: 65, the H-FR4 of the antigen-binding protein has an amino acid substitution (e.g., conservative amino acid substitution, etc.) at one or more amino acids selected from the group consisting of X 5 and X 6 .

[0163] WGQGX5 X 6 VTVSS (SEQ ID NO: 65) (where X 5 may be T or V, and X 6 may be L or M)

[0164] In the present application, the H-FR4 of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 20 and SEQ ID NO: 30.

[0165] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 62, the H-FR2 may contain the amino acid sequence represented by SEQ ID NO: 63, the H-FR3 may contain the amino acid sequence represented by SEQ ID NO: 64, and the H-FR4 may contain the amino acid sequence represented by SEQ ID NO: 65.

[0166] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 14 and SEQ ID NO: 27, the H-FR2 may contain the amino acid sequence represented by any one of SEQ ID NO: 16 and SEQ ID NO: 28, the H-FR3 may contain the amino acid sequence represented by any one of SEQ ID NO: 18 and SEQ ID NO: 29, and the H-FR4 may contain the amino acid sequence represented by any one of SEQ ID NO: 20 and SEQ ID NO: 30.

[0167] In the present application, the H-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 14, the H-FR2 may include the amino acid sequence represented by SEQ ID NO: 16, the H-FR3 may include the amino acid sequence represented by SEQ ID NO: 18, and the H-FR4 may include the amino acid sequence represented by SEQ ID NO: 20. For example, the antigen-binding protein may include antibody 50A6 or an antigen-binding fragment having the same H-FR1-4 as this.

[0168] In the present application, the H-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 27, the H-FR2 may include the amino acid sequence represented by SEQ ID NO: 28, the H-FR3 may include the amino acid sequence represented by SEQ ID NO: 29, and the H-FR4 may include the amino acid sequence represented by SEQ ID NO: 30. For example, the antigen-binding protein may include antibody JYB1931A63 or an antigen-binding fragment having the same H-FR1-4 as this.

[0169] In the present application, the antigen-binding protein may include a heavy chain variable region, and the heavy chain variable region may include the amino acid sequence represented by SEQ ID NO: 67. For example, the antigen-binding protein includes VH, and the VH has an amino acid substitution (for example, a conservative amino acid substitution, etc.) for one or more amino acids selected from the group consisting of X 16 、X 19 、X 40 、X 42 、X 73 、X 77 、X 78 、X 84 、X 88 、X 93 、X 119 、X 120 from the group consisting of X

[0170] EVQLVESGGGLVQPGX 16 SLX 19LSCAASGFTFNDYNMAWVRQX 40 PX 42 KGLEWVATILFDGSRTYYRDSVKGRFTISRX 73 NAKX 77 X 78 LYLQMX 84 SLRX 88 EDTAX 93 YYCSTESPYYSEGYYQGYFDYWGQGX 119 X 120 VTVSS(SEQ ID NO: 67)(where X 16 may be G or R, X 19 may be R or S, X 40 may be A or T, X 42 may be G or K, X 73 may be D or E, X 77 may be N or S, X 78 may be S or T, X 84 may be D or N, X 88 may be A or S, X 93 may be T or V, X 119 may be T or V, X 120 may be L or M)

[0171] In the present application, the heavy chain variable region of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 13 and SEQ ID NO: 26.

[0172] In the present application, the antigen-binding protein may contain a heavy chain constant region, and the heavy chain constant region may contain a constant region derived from IgG or a constant region derived from IgY.

[0173] For example, the heavy chain constant region of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 56.

[0174] In the present application, the antigen-binding protein may include at least one CDR within the variable region VL of the antibody light chain, and the VL may include the amino acid sequence represented by SEQ ID NO: 66 or SEQ ID NO: 76.

[0175] For example, the VL may include the amino acid sequence represented by either SEQ ID NO: 66 or SEQ ID NO: 76. In the present application, the LCDRs of the isolated antigen-binding protein may be divided in any manner, and as long as the VL is homologous to the amino acid sequence represented by either SEQ ID NO: 66 or SEQ ID NO: 76, the LCDRs divided in any manner are within the scope of the patent of the present application.

[0176] In the present application, the antigen-binding protein may include the variable region VL of the light chain, and the VL may include at least one, at least two, or at least three of LCDR1, LCDR2, and LCDR3.

[0177] In the present application, the LCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 11. For example, the LCDR3 of the antigen-binding protein may be defined by the Chothia numbering system.

[0178] In the present application, the LCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 9. For example, the LCDR2 of the antigen-binding protein may be defined by the Chothia numbering system.

[0179] In the present application, the LCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 7. For example, the LCDR1 of the antigen-binding protein may be defined by the Chothia numbering system.

[0180] For example, the LCDR1 of the antigen-binding protein of the present application may contain the amino acid sequence shown in SEQ ID NO: 7, the LCDR2 may contain the amino acid sequence shown in SEQ ID NO: 9, and the LCDR3 may contain the amino acid sequence shown in SEQ ID NO: 11. For example, the antigen-binding protein may contain antibody 50A6, JYB1931A63, or an antigen-binding fragment having the same LCDR3 (for example, the same LCDR1-3) as this.

[0181] For example, the VL of the antigen-binding protein may contain framework regions L-FR1, L-FR2, L-FR3, and L-FR4.

[0182] In the present application, the L-FR1 of the antigen-binding protein may contain the amino acid sequence shown in SEQ ID NO: 58. For example, compared with the sequence shown in SEQ ID NO: 58, the L-FR1 of the antigen-binding protein has amino acid substitutions (for example, conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of X 1 , X 13 , X 14 , X 18 , X 19 and X 20 .

[0183] X 1 IVLTQSPATLSX 13 X 14 PGEX 18 X 19 X 20 LSC (SEQ ID NO: 58) (where X 1 may be E or N, X 13 may be L or V, X 14 may be S or T, X 18 may be R or S, X 19 may be A or V, X 20 may be S or T)

[0184] In the present application, the L-FR1 of the antigen-binding protein may contain the amino acid sequence represented by any of SEQ ID NO: 6 and SEQ ID NO: 22.

[0185] In the present application, the L-FR2 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 59. For example, the L-FR2 of the antigen-binding protein has X 5 , X 6 , X 7 , X 8 and X 9 wherein one or more amino acid substitutions (such as conservative amino acid substitutions, etc.) are present among the amino acids selected from the group consisting of

[0186] WYQQX 5 X 6 X 7 X 8 X 9 PRLLIK (SEQ ID NO: 59) (where X 5 may be K or R, X 6 may be P or S, X 7 may be G or N, X 8 may be E or Q, X 9 may be A or S)

[0187] In the present application, the L-FR2 of the antigen-binding protein may contain the amino acid sequence represented by any of SEQ ID NO: 8 and SEQ ID NO: 23.

[0188] In the present application, the L-FR3 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 60. For example, the L-FR3 of the antigen-binding protein has X 4 , X 18 , X 20 , X 21 , X 22 , X 24 , X 28 and X29 There are amino acid substitutions (such as conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of

[0189] GIPX 4 RFSGSGSGTDFTLX 18 IX 20 X 21 X 22 EX 24 EDFX 28 X 29 YYC (SEQ ID NO: 60) (where X 4 may be A or S, X 18 may be S or T, X 20 may be N or S, X 21 may be R or S, X 22 may be L or V, X 24 may be P or S, X 28 may be A or S, X 29 may be I or V)

[0190] In the present application, the L-FR3 of the antigen-binding protein may include an amino acid sequence represented by any of SEQ ID NO: 10 and SEQ ID NO: 24.

[0191] In the present application, the L-FR4 of the antigen-binding protein may include an amino acid sequence represented by SEQ ID NO: 61. For example, compared with the sequence represented by SEQ ID NO: 61, the L-FR4 of the antigen-binding protein has amino acid substitutions (such as conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of X 3 X 9 and X 10

[0192] FGX 3 GTKLEX 9 X 10 (SEQ ID NO: 61) (where X 3 may be A or Q, X 9 ​may be I or L, X 10 may be K or R)

[0193] In the present application, the L-FR4 of the antigen-binding protein may include the amino acid sequence represented by any one of SEQ ID NO: 12 and SEQ ID NO: 25.

[0194] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 58, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 59, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 60, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 61.

[0195] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by any one of SEQ ID NO: 6 and SEQ ID NO: 22, the L-FR2 may include the amino acid sequence represented by any one of SEQ ID NO: 8 and SEQ ID NO: 23, the L-FR3 may include the amino acid sequence represented by any one of SEQ ID NO: 10 and SEQ ID NO: 24, and the L-FR4 may include the amino acid sequence represented by any one of SEQ ID NO: 12 and SEQ ID NO: 25.

[0196] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 6, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 8, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 10, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 12. For example, the antigen-binding protein may include antibody 50A6 or an antibody having the same L-FR1-4 as this.

[0197] In the present application, the L-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 22, the L-FR2 may contain the amino acid sequence represented by SEQ ID NO: 23, the L-FR3 may contain the amino acid sequence represented by SEQ ID NO: 24, and the L-FR4 may contain the amino acid sequence represented by SEQ ID NO: 25. For example, the antigen-binding protein may include antibody JYB1931A63 or an antibody having the same H-FR1-4 as it.

[0198] In the present application, the antigen-binding protein may contain a light chain variable region VL, and the VL may contain the amino acid sequence represented by SEQ ID NO: 66. For example, the VL of the antigen-binding protein has 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 One or more amino acid substitutions (e.g., conservative amino acid substitutions, etc.) exist in one or more amino acids selected from the group consisting of 1

[0199] X 1 IVLTQSPATLSX 13 X 14 PGEX 18 X 19 X 20 LSCRASQSVSTSIHWYQQX 39 X 40 X 41 X 42 X 43PRLLIKYASRSISGIPX 60 RFSGSGSGTDFTLX 74 IX 76 X 77 X 78 EX 80 EDFX 84 X 85 YYCQQSYSSLYTFGX 100 GTKLEX 106 X 107 (SEQ ID NO: 66)(where X 1 may be E or N, X 13 may be L or V, X 14 may be S or T, X 18 may be R or S, X 19 may be A or V, X 20 may be S or T, X 39 may be K or R, X 40 may be P or S, X 41 may be G or N, X 42 may be E or Q, X 43 may be A or S, X 60 may be A or S, X 74 may be S or T, X 76 may be N or S, X 77 may be R or S, X 78 may be L or V, X 80 may be P or S, X 84 may be A or S, X 85 may be I or V, X 100 may be A or Q, X 106 may be I or L, X 107 may be K or R)

[0200] In the present application, the variable region of the light chain of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 5 and SEQ ID NO: 21.

[0201] In the present application, the antigen-binding protein may include a light-chain constant region, and the light-chain constant region may include a constant region derived from Igκ or a constant region derived from Igλ.

[0202] For example, the light-chain constant region may include a constant region derived from Igκ.

[0203] For example, the light-chain constant region of the antigen-binding protein includes the amino acid sequence represented by SEQ ID NO: 57.

[0204] In the present application, the antigen-binding protein may include HCDR1-3 and LCDR1-3. For example, HCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 15, HCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 17, HCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 19, LCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 7, LCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 9, and LCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 11. For example, the antigen-binding protein may include antibody 50A6, JYB1931A63, or an antigen-binding fragment having the same HCDR3 (for example, the same HCDR1-3) and LCDR3 (for example, the same LCDR1-3) as these.

[0205] In the present application, the antigen-binding protein may include a heavy-chain variable region and a light-chain variable region. The heavy-chain variable region of the antigen-binding protein may include HCDR1-3 and H-FR1-4. The light-chain variable region of the antigen-binding protein may include LCDR1-3 and L-FR1-4. For example, the HCDR1 may include the amino acid sequence represented by SEQ ID NO: 15, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 17, the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 19, the LCDR1 may include the amino acid sequence represented by SEQ ID NO: 7, the LCDR2 may include the amino acid sequence represented by SEQ ID NO: 9, and the LCDR3 may include the amino acid sequence represented by SEQ ID NO: 11. For example, the H-FR1 may include the amino acid sequence represented by SEQ ID NO: 14, the H-FR2 may include the amino acid sequence represented by SEQ ID NO: 16, the H-FR3 may include the amino acid sequence represented by SEQ ID NO: 18, the H-FR4 may include the amino acid sequence represented by SEQ ID NO: 20, the L-FR1 may include the amino acid sequence represented by SEQ ID NO: 6, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 8, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 10, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 12. For example, the heavy-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 13. For example, the antigen-binding protein may include antibody 50A6 or an antigen-binding protein having the same heavy-chain variable region as this. For example, the light-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 5. For example, the antigen-binding protein may include antibody 50A6 or an antigen-binding protein having the same light-chain variable region as this.

[0206] In the present application, the antigen-binding protein may include a heavy-chain variable region and a light-chain variable region. The heavy-chain variable region may include HCDR1-3 and H-FR1-4. The light-chain variable region may include LCDR1-3 and L-FR1-4. For example, the HCDR1 may include the amino acid sequence represented by SEQ ID NO: 15, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 17, the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 19, the LCDR1 may include the amino acid sequence represented by SEQ ID NO: 7, the LCDR2 may include the amino acid sequence represented by SEQ ID NO: 9, and the LCDR3 may include the amino acid sequence represented by SEQ ID NO: 11. For example, the H-FR1 may include the amino acid sequence represented by SEQ ID NO: 27, the H-FR2 may include the amino acid sequence represented by SEQ ID NO: 28, the H-FR3 may include the amino acid sequence represented by SEQ ID NO: 29, the H-FR4 may include the amino acid sequence represented by SEQ ID NO: 30, the L-FR1 may include the amino acid sequence represented by SEQ ID NO: 22, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 23, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 24, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 25. For example, the heavy-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 26. For example, the antigen-binding protein may include antibody JYB1931A63 or an antigen-binding protein having the same heavy-chain variable region as this. For example, the light-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 21. For example, the antigen-binding protein may include antibody JYB1931A63 or an antigen-binding protein having the same light-chain variable region as this.

[0207] In the present application, the antigen-binding protein may include a heavy-chain variable region VH, and the VH may include at least one, two, or three of HCDR1, HCDR2, and HCDR3.

[0208] In the present application, the HCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 45. For example, the HCDR3 sequence of the antigen-binding protein may be defined by the Chothia numbering system.

[0209] In the present application, the HCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 43. For example, the HCDR2 sequence of the antigen-binding protein may be defined by the Chothia numbering system.

[0210] In the present application, the HCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 41. For example, the HCDR1 sequence of the antigen-binding protein may be defined by the Chothia numbering system.

[0211] For example, the HCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 41, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 43, and the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 45. For example, the antigen-binding protein may include antibody 47A1, JYB1931A13, or an antigen-binding fragment having the same HCDR3 (for example, the same HCDR1-3) as this.

[0212] For example, the VH of the antigen-binding protein may include framework regions H-FR1, H-FR2, H-FR3, and H-FR4.

[0213] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 72. For example, compared with the sequence represented by SEQ ID NO: 72, the H-FR1 of the antigen-binding protein has an amino acid substitution (e.g., conservative amino acid substitution, etc.) at one or more amino acids selected from the group consisting of X 1 X 17 and X 25 .

[0214] X 1 VQLQESGPGLVKPSQX 17 LSLTCTVX 25 (SEQ ID NO: 72) (where X 1 may be D or Q, X 17 may be S or T, and X 25 may be S or T)

[0215] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 40 and SEQ ID NO: 53.

[0216] In the present application, the H-FR2 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 73. For example, compared with the sequence represented by SEQ ID NO: 73, the H-FR2 of the antigen-binding protein has an amino acid substitution (e.g., conservative amino acid substitution, etc.) at one or more amino acids selected from the group consisting of X 8 X 11 X 12 and X 16 .

[0217] AWNWIRQX 8 PGX 11 X 12 LEWX 16 GYI(SEQ ID NO: 73) (where X 8 may be F or P, X 11 may be K or N, X 12 may be G or K, X16 may be I or M)

[0218] In the present application, the H-FR2 of the antigen-binding protein may include an amino acid sequence represented by any of SEQ ID NO: 42 and SEQ ID NO: 54.

[0219] In the present application, the H-FR3 of the antigen-binding protein may include an amino acid sequence represented by SEQ ID NO: 74. For example, the H-FR3 of the antigen-binding protein, compared with the sequence represented by SEQ ID NO: 74, has 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 and has one or more amino acid substitutions (for example, conservative amino acid substitutions, etc.) in the amino acids selected from the group consisting of X

[0220] TSYNPSLKSRX 11 X 12 IX 14 RX 16 TSX 19 NQFX 23 LX 25 LX 27 SVTX 31 X 32 DTAX 36 YX 38 CAR (SEQ ID NO: 74) (where X 11 may be I or V, X 12 may be S or T, X 14 may be S or T, X 16 may be D or N, X 19 may be K or T, X 23 may be F or S, X 25 may be K or Q, X 27 may be N or S, X31 may be A or T, X 32 may be A or E, X 36 may be T or V, X 38 may be F or Y)

[0221] In the present application, the H-FR3 of the antigen-binding protein may contain an amino acid sequence represented by any one of SEQ ID NO: 44 and SEQ ID NO: 55.

[0222] In the present application, the H-FR4 of the antigen-binding protein may contain an amino acid sequence represented by SEQ ID NO: 75. For example, the H-FR4 of the antigen-binding protein, compared with the sequence represented by SEQ ID NO: 75, X 6 and X 7 has one or more amino acid substitutions (e.g., conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of.

[0223] WGQGTX 6 X 7 TVSS (SEQ ID NO: 75) (where X 6 may be L or T, X 7 may be L or V)

[0224] In the present application, the H-FR4 of the antigen-binding protein may contain an amino acid sequence represented by any one of SEQ ID NO: 46 and SEQ ID NO: 30.

[0225] In the present application, the H-FR1 of the antigen-binding protein may contain an amino acid sequence represented by SEQ ID NO: 72, the H-FR2 may contain an amino acid sequence represented by SEQ ID NO: 73, the H-FR3 may contain an amino acid sequence represented by SEQ ID NO: 74, and the H-FR4 may contain an amino acid sequence represented by SEQ ID NO: 75.

[0226] In the present application, the H-FR1 of the antigen-binding protein may contain an amino acid sequence represented by any one of SEQ ID NO: 40 and SEQ ID NO: 53, the H-FR2 may contain an amino acid sequence represented by any one of SEQ ID NO: 42 and SEQ ID NO: 54, the H-FR3 may contain an amino acid sequence represented by any one of SEQ ID NO: 44 and SEQ ID NO: 55, and the H-FR4 may contain an amino acid sequence represented by any one of SEQ ID NO: 46 and SEQ ID NO: 30.

[0227] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 40, the H-FR2 may contain the amino acid sequence represented by SEQ ID NO: 42, the H-FR3 may contain the amino acid sequence represented by SEQ ID NO: 44, and the H-FR4 may contain the amino acid sequence represented by SEQ ID NO: 46. For example, the antigen-binding protein may contain antibody 47A1 or an antigen-binding fragment having the same H-FR1-4 as this.

[0228] In the present application, the H-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 53, the H-FR2 may contain the amino acid sequence represented by SEQ ID NO: 54, the H-FR3 may contain the amino acid sequence represented by SEQ ID NO: 55, and the H-FR4 may contain the amino acid sequence represented by SEQ ID NO: 30. For example, the antigen-binding protein may contain antibody JYB1931A13 or an antigen-binding fragment having the same H-FR1-4 as this.

[0229] In the present application, the antigen-binding protein may contain a heavy chain variable region, and the heavy chain variable region may contain the amino acid sequence represented by SEQ ID NO: 77. For example, the antigen-binding protein contains VH, and the VH, compared with the sequence represented by SEQ ID NO: 77, is X1 , 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 and X 110 There are amino acid substitutions (e.g., conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of

[0230] X 1 VQLQESGPGLVKPSQX 17 LSLTCTVX 25 GYSITSDYAWNWIRQX 41 PGX 44 X 45 LEWX 49 GYISYSGRTSYNPSLKSRX 68 X 69 IX 71 RX 73 TSX 76 NQFX 80 LX 82 LX 84 SVTX 88 X 89 DTAX 93 YX 95 CARYWGDYWGQGTX 109 X 110 TVSS (SEQ ID NO: 77) (where X 1 may be D or Q, X 17 may be S or T, X 25 may be S or T, X 41 may be F or P, X 44 may be K or N, X 45 may be G or K, X 49may be I or M, X 68 may be I or V, X 69 may be S or T, X 71 may be S or T, X 73 may be D or N, X 76 may be K or T, X 80 may be F or S, X 82 may be K or Q, X 84 may be N or S, X 88 may be A or T, X 89 may be A or E, X 93 may be T or V, X 95 may be F or Y, X 109 may be L or T, X 110 may be L or V)

[0231] In the present application, the heavy chain variable region of the antigen-binding protein may contain the amino acid sequence represented by any one of SEQ ID NO: 39 and SEQ ID NO: 52.

[0232] In the present application, the antigen-binding protein may contain a heavy chain constant region, and the heavy chain constant region may contain a constant region derived from IgG or a constant region derived from IgY.

[0233] For example, the heavy chain constant region of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 56.

[0234] In the present application, the antigen-binding protein may contain a light chain variable region VL, and the VL may contain at least one, at least two, or at least three of LCDR1, LCDR2, and LCDR3.

[0235] In the present application, the LCDR3 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 37. For example, the LCDR3 of the antigen-binding protein may be defined by the Chothia numbering system.

[0236] In the present application, the LCDR2 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 35. For example, the LCDR2 of the antigen-binding protein may be defined by the Chothia numbering system.

[0237] In the present application, the LCDR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 33. For example, the LCDR1 of the antigen-binding protein may be defined by the Chothia numbering system.

[0238] For example, the LCDR1 of the antigen-binding protein in the present application may contain the amino acid sequence represented by SEQ ID NO: 33, the LCDR2 may contain the amino acid sequence represented by SEQ ID NO: 35, and the LCDR3 may contain the amino acid sequence represented by SEQ ID NO: 37. For example, the antigen-binding protein may contain antibody 47A1, JYB1931A13, or an antigen-binding fragment having the same LCDR3 (for example, the same LCDR1-3).

[0239] For example, the VL of the antigen-binding protein may contain framework regions L-FR1, L-FR2, L-FR3, and L-FR4.

[0240] In the present application, the L-FR1 of the antigen-binding protein may contain the amino acid sequence represented by SEQ ID NO: 68. For example, the L-FR1 of the antigen-binding protein, compared with the sequence represented by SEQ ID NO: 68, is X 3 、X 8 、X 9 、X 10 、X 11 、X13 , X 20 and X 21 There are amino acid substitutions (e.g., conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of

[0241] DIX 3 MTQSX 8 X 9 X 10 X 11 SX 13 SVGDRVX 20 X 21 TC (SEQ ID NO: 68) (where X 3 may be Q or V, X 8 may be P or Q, X 9 may be K or S, X 10 may be F or S, X 11 may be L or M, X 13 may be A or T, X 20 may be S or T, X 21 may be I or V)

[0242] In the present application, the L-FR1 of the antigen-binding protein may contain an amino acid sequence represented by any of SEQ ID NO: 32 and SEQ ID NO: 48.

[0243] In the present application, the L-FR2 of the antigen-binding protein may contain an amino acid sequence represented by SEQ ID NO: 69. For example, the L-FR2 of the antigen-binding protein, compared with the sequence represented by SEQ ID NO: 69, has X 8 and X 9 There are amino acid substitutions (e.g., conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of

[0244] WFQQKPGX 8 X 9 PKPLIY (SEQ ID NO: 69) (where X 8 may be K or Q, X 9It may be A or S)

[0245] In the present application, the L-FR2 of the antigen-binding protein may contain an amino acid sequence represented by any of SEQ ID NO: 34 and SEQ ID NO: 49.

[0246] In the present application, the L-FR3 of the antigen-binding protein may contain an amino acid sequence represented by SEQ ID NO: 70. For example, the L-FR3 of the antigen-binding protein has X 4 , X 7 , X 21 , X 22 , X 24 , X 27 and X 29 One or more amino acid substitutions (such as conservative amino acid substitutions, etc.) are present in one or more amino acids selected from the group consisting of.

[0247] GVPX 4 RFX 7 GSGSGTDFTLTISX 21 X 22 QX 24 EDX 27 AX 29 YFC (SEQ ID NO: 70) (where X 4 may be D or S, X 7 may be S or T, X 21 may be N or S, X 22 may be L or V, X 24 may be P or S, X 27 may be F or L, X 29 may be E or T)

[0248] In the present application, the L-FR3 of the antigen-binding protein may contain an amino acid sequence represented by any of SEQ ID NO: 36 and SEQ ID NO: 50.

[0249] In the present application, the L-FR4 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 71. For example, the L-FR4 of the antigen-binding protein may have amino acid substitutions (such as conservative amino acid substitutions, etc.) at one or more amino acids selected from the group consisting of X 3 、X 7 、X 9 、X 10 .

[0250] FGX 3 GTKX 7 EX 9 X 10 (SEQ ID NO: 71) (where X 3 may be A or G, X 7 may be L or V, X 9 may be I or L, X 10 may be K or N)

[0251] In the present application, the L-FR4 of the antigen-binding protein may include the amino acid sequence represented by any one of SEQ ID NO: 38 and SEQ ID NO: 51.

[0252] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 68, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 69, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 70, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 71.

[0253] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by any one of SEQ ID NO: 32 and SEQ ID NO: 48, the L-FR2 may include the amino acid sequence represented by any one of SEQ ID NO: 34 and SEQ ID NO: 49, the L-FR3 may include the amino acid sequence represented by any one of SEQ ID NO: 36 and SEQ ID NO: 50, and the L-FR4 may include the amino acid sequence represented by any one of SEQ ID NO: 38 and SEQ ID NO: 51.

[0254] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 32, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 34, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 36, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 38. For example, the antigen-binding protein may include antibody 47A1 or an antibody having the same L-FR1-4 as this.

[0255] In the present application, the L-FR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 48, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 49, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 50, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 51. For example, the antigen-binding protein may include antibody JYB1931A13 or an antibody having the same H-FR1-4 as this.

[0256] In the present application, the antigen-binding protein may include a light chain variable region VL, and the VL may include the amino acid sequence represented by SEQ ID NO: 76. For example, the VL of the antigen-binding protein, compared with the sequence represented by SEQ ID NO: 76, is 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 There are one or more amino acid substitutions (e.g., conservative amino acid substitutions, etc.) in one or more amino acids selected from the group consisting of

[0257] DIX 3 MTQSX 8 , X 9 , X 10 , X 11 SX 13 SVGDRVX 20 , X 21 TCKASQNVGSNVAWFQQKPGX 42 , X 43 PKPLIYSASYRYSGVPX 60 RFX 63 GSGSGTDFTLTISX 77 , X 78 QX 80 EDX 83 , AX 85 YFCHQYNTYPLTFGX 100 GTK X 104 , EX 106 , X 107 (SEQ ID NO: 76) (where X 3 may be Q or V, X 8 may be P or Q, X 9 may be K or S, X 10 may be F or S, X 11 may be L or M, X 13 may be A or T, X 20 may be S or T, X 21 may be I or V, X42 may be K or Q, X 43 may be A or S, X 60 may be D or S, X 63 may be S or T, X 77 may be N or S, X 78 may be L or V, X 80 may be P or S, X 83 may be F or L, X 85 may be E or T, X 100 may be A or G, X 104 may be L or V, X 106 may be I or L, X 107 may be K or N) In the present application, the light chain variable region of the antigen-binding protein may contain the amino acid sequence represented by any of SEQ ID NO: 31 and 47.

[0258] In the present application, the antigen-binding protein may contain a light chain constant region, and the light chain constant region may contain a constant region derived from Igκ or a constant region derived from Igλ.

[0259] For example, the light chain constant region may contain a constant region derived from Igκ.

[0260] For example, the light chain constant region of the antigen-binding protein contains the amino acid sequence represented by SEQ ID NO: 57.

[0261] In the present application, the antigen-binding protein may include HCDR1-3 and LCDR1-3. For example, the HCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 41, the HCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 43, the HCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 45, the LCDR1 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 33, the LCDR2 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 35, and the LCDR3 of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 37. For example, the antigen-binding protein may include antibody 47A1, JYB1931A13, or an antigen-binding fragment having the same HCDR3 (e.g., the same HCDR1-3) and LCDR3 (e.g., the same LCDR1-3) as these.

[0262] In the present application, the antigen-binding protein may include a heavy-chain variable region and a light-chain variable region. The heavy-chain variable region of the antigen-binding protein may include HCDR1-3 and H-FR1-4. The light-chain variable region of the antigen-binding protein may include LCDR1-3 and L-FR1-4. For example, the HCDR1 may include the amino acid sequence represented by SEQ ID NO: 41, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 43, the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 45, the LCDR1 may include the amino acid sequence represented by SEQ ID NO: 33, the LCDR2 may include the amino acid sequence represented by SEQ ID NO: 35, and the LCDR3 may include the amino acid sequence represented by SEQ ID NO: 37. For example, the H-FR1 may include the amino acid sequence represented by SEQ ID NO: 40, the H-FR2 may include the amino acid sequence represented by SEQ ID NO: 42, the H-FR3 may include the amino acid sequence represented by SEQ ID NO: 44, the H-FR4 may include the amino acid sequence represented by SEQ ID NO: 46, the L-FR1 may include the amino acid sequence represented by SEQ ID NO: 32, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 34, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 36, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 38. For example, the heavy-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 39. For example, the antigen-binding protein may include antibody 47A1 or an antigen-binding protein having the same heavy-chain variable region as this. For example, the light-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 31. For example, the antigen-binding protein may include antibody 47A1 or an antigen-binding protein having the same light-chain variable region as this.

[0263] In the present application, the antigen-binding protein may include a heavy-chain variable region and a light-chain variable region. The heavy-chain variable region may include HCDR1-3 and H-FR1-4. The light-chain variable region may include LCDR1-3 and L-FR1-4. For example, the HCDR1 may include the amino acid sequence represented by SEQ ID NO: 41, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 43, the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 45, the LCDR1 may include the amino acid sequence represented by SEQ ID NO: 33, the LCDR2 may include the amino acid sequence represented by SEQ ID NO: 35, and the LCDR3 may include the amino acid sequence represented by SEQ ID NO: 37. For example, the H-FR1 may include the amino acid sequence represented by SEQ ID NO: 53, the H-FR2 may include the amino acid sequence represented by SEQ ID NO: 54, the H-FR3 may include the amino acid sequence represented by SEQ ID NO: 55, the H-FR4 may include the amino acid sequence represented by SEQ ID NO: 30, the L-FR1 may include the amino acid sequence represented by SEQ ID NO: 48, the L-FR2 may include the amino acid sequence represented by SEQ ID NO: 49, the L-FR3 may include the amino acid sequence represented by SEQ ID NO: 50, and the L-FR4 may include the amino acid sequence represented by SEQ ID NO: 51. For example, the heavy-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 52. For example, the antigen-binding protein may include antibody JYB1931A13 or an antigen-binding protein having the same heavy-chain variable region as this. For example, the light-chain variable region of the antigen-binding protein may include the amino acid sequence represented by SEQ ID NO: 47. For example, the antigen-binding protein may include antibody JYB1931A13 or an antigen-binding protein having the same light-chain variable region as this.

[0264] In the present application, the isolated antigen-binding protein may competitively bind to a reference antibody and the human MASP-2 protein, the reference antibody may include a heavy chain variable region VH, and the VH may include at least one, two, or three of HCDR1, HCDR2, and HCDR3.

[0265] In the present application, the HCDR3 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 19. For example, the sequence of the HCDR3 of the reference antibody may be defined by the Chothia numbering system.

[0266] In the present application, the HCDR2 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 17. For example, the sequence of the HCDR2 of the reference antibody may be defined by the Chothia numbering system.

[0267] In the present application, the HCDR1 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 15. For example, the sequence of the HCDR1 of the reference antibody may be defined by the Chothia numbering system.

[0268] For example, the HCDR1 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 15, the HCDR2 may include the amino acid sequence shown in SEQ ID NO: 17, and the HCDR3 may include the amino acid sequence shown in SEQ ID NO: 19. For example, the reference antibody may include antibody 50A6, JYB1931A63, or an antigen-binding protein having the same HCDR3 (e.g., the same HCDR1-3) as this.

[0269] In the present application, the reference antibody may include a heavy chain variable region, and the heavy chain variable region may include the amino acid sequence shown in SEQ ID NO: 67.

[0270] EVQLVESGGGLVQPGX 16 SLX19 LSCAASGFTFNDYNMAWVRQX 40 PX 42 KGLEWVATILFDGSRTYYRDSVKGRFTISRX 73 NAKX 77 X 78 LYLQMX 84 SLRX 88 EDTAX 93 YYCSTESPYYSEGYYQGYFDYWGQGX 119 X 120 VTVSS(SEQ ID NO: 67)(where X 16 may be G or R, X 19 may be R or S, X 40 may be A or T, X 42 may be G or K, X 73 may be D or E, X 77 may be N or S, X 78 may be S or T, X 84 may be D or N, X 88 may be A or S, X 93 may be T or V, X 119 may be T or V, X 120 may be L or M)

[0271] In the present application, the heavy chain variable region of the reference antibody may contain an amino acid sequence represented by any of SEQ ID NO: 13 and SEQ ID NO: 26.

[0272] In the present application, the reference antibody may contain a heavy chain constant region, and the heavy chain constant region may contain a constant region derived from IgG or a constant region derived from IgY.

[0273] For example, the heavy chain constant region of the reference antibody may contain an amino acid sequence represented by SEQ ID NO: 56.

[0274] In the present application, the reference antibody may include a light chain variable region VL, and the VL may include LCDR1, LCDR2, and LCDR3.

[0275] In the present application, the LCDR3 of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 11. For example, the sequence of the LCDR3 of the reference antibody may be defined by the Chothia numbering system.

[0276] In the present application, the LCDR2 of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 9. For example, the sequence of the LCDR2 of the reference antibody may be defined by the Chothia numbering system.

[0277] In the present application, the LCDR1 of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 7. For example, the sequence of the LCDR1 of the reference antibody may be defined by the Chothia numbering system.

[0278] For example, the LCDR1 of the reference antibody in the present application may include the amino acid sequence represented by SEQ ID NO: 7, the LCDR2 may include the amino acid sequence represented by SEQ ID NO: 9, and the LCDR3 may include the amino acid sequence represented by SEQ ID NO: 11. For example, the reference antibody may include antibody 50A6, JYB1931A63, or an antigen-binding protein having the same LCDR3 (for example, the same LCDR1-3) as this.

[0279] In the present application, the reference antibody may include a light chain variable region, and the light chain variable region may include the amino acid sequence represented by SEQ ID NO: 66.

[0280] X 1 IVLTQSPATLSX 13 X 14 PGEX 18 X 19 X 20LSCRASQSVSTSIHWYQQX 39 X 40 X 41 X 42 X 43 PRLLIKYASRSISGIPX 60 RFSGSGSGTDFTLX 74 IX 76 X 77 X 78 EX 80 EDFX 84 X 85 YYCQQSYSSLYTFGX 100 GTKLEX 106 X 107 (SEQ ID NO: 66)(where X 1 may be E or N, X 13 may be L or V, X 14 may be S or T, X 18 may be R or S, X 19 may be A or V, X 20 may be S or T, X 39 may be K or R, X 40 may be P or S, X 41 may be G or N, X 42 may be E or Q, X 43 may be A or S, X 60 may be A or S, X 74 may be S or T, X 76 may be N or S, X 77 may be R or S, X 78 may be L or V, X 80 may be P or S, X 84 may be A or S, X 85 may be I or V, X 100 may be A or Q, X 106 may be I or L, X 107 may be K or R)

[0281] In the present application, the light chain variable region of the reference antibody may include an amino acid sequence represented by any one of SEQ ID NO: 5 and SEQ ID NO: 21.

[0282] In the present application, the reference antibody may include HCDR1-3 and LCDR1-3. For example, the HCDR1 may include an amino acid sequence represented by SEQ ID NO: 15, the HCDR2 may include an amino acid sequence represented by SEQ ID NO: 17, the HCDR3 may include an amino acid sequence represented by SEQ ID NO: 19, the LCDR1 may include an amino acid sequence represented by SEQ ID NO: 7, the LCDR2 may include an amino acid sequence represented by SEQ ID NO: 9, and the LCDR3 may include an amino acid sequence represented by SEQ ID NO: 11. For example, the reference antibody may include antibody 50A6, JYB1931A63, or an antigen-binding protein having the same HCDR3 (for example, the same HCDR1-3) and LCDR3 (for example, the same LCDR1-3) as these.

[0283] In the present application, the isolated antigen-binding protein may also competitively bind to the reference antibody and the human MASP-2 protein. The reference antibody may include a heavy chain variable region VH, and the VH may include at least one, two, or three of HCDR1, HCDR2, and HCDR3.

[0284] In the present application, the HCDR3 of the reference antibody may include an amino acid sequence represented by SEQ ID NO: 45. For example, the sequence of the HCDR3 of the reference antibody may be defined by the Chothia numbering system.

[0285] In the present application, the HCDR2 of the reference antibody may include an amino acid sequence represented by SEQ ID NO: 43. For example, the sequence of the HCDR2 of the reference antibody may be defined by the Chothia numbering system.

[0286] In the present application, the HCDR1 of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 41. For example, the sequence of the HCDR1 of the reference antibody may be defined by the Chothia numbering system.

[0287] For example, the HCDR1 of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 41, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 43, and the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 45. For example, the reference antibody may include antibody 47A1, JYB1931A13, or an antigen-binding protein having the same HCDR3 (e.g., the same HCDR1-3) as this.

[0288] In the present application, the reference antibody may include a heavy chain variable region, and the heavy chain variable region may include the amino acid sequence represented by SEQ ID NO: 77.

[0289] X 1 VQLQESGPGLVKPSQ X 17 LSLTCTV X 25 GYSITSDYAWNWIRQX 41 PGX 44 X 45 LEWX 49 GYISYSGRTSYNPSLKSRX 68 X 69 IX 71 RX 73 TSX 76 NQFX 80 LX 82 LX 84 SVTX 88 X 89 DTAX 93 YX 95 CARYWGDYWGQGTX 109 X 110 TVSS(SEQ ID NO: 77)(where X 1 may be D or Q, and X 17 may be S or T, and X 25may be S or T, X 41 may be F or P, X 44 may be K or N, X 45 may be G or K, X 49 may be I or M, X 68 may be I or V, X 69 may be S or T, X 71 may be S or T, X 73 may be D or N, X 76 may be K or T, X 80 may be F or S, X 82 may be K or Q, X 84 may be N or S, X 88 may be A or T, X 89 may be A or E, X 93 may be T or V, X 95 may be F or Y, X 109 may be L or T, X 110 may be L or V.

[0290] In the present application, the heavy chain variable region of the reference antibody may contain an amino acid sequence represented by any of SEQ ID NO: 39 and SEQ ID NO: 52.

[0291] In the present application, the reference antibody may contain a heavy chain constant region, and the heavy chain constant region may contain a constant region derived from IgG or a constant region derived from IgY.

[0292] For example, the heavy chain constant region of the reference antibody may contain an amino acid sequence represented by SEQ ID NO: 56.

[0293] In the present application, the reference antibody may contain a light chain variable region VL, and the VL may contain LCDR1, LCDR2, and LCDR3.

[0294] In the present application, the LCDR3 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 37. For example, the sequence of the LCDR3 of the reference antibody may be defined by the Chothia numbering system.

[0295] In the present application, the LCDR2 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 35. For example, the sequence of the LCDR2 of the reference antibody may be defined by the Chothia numbering system.

[0296] In the present application, the LCDR1 of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 33. For example, the sequence of the LCDR1 of the reference antibody may be defined by the Chothia numbering system.

[0297] For example, the LCDR1 of the reference antibody in the present application may include the amino acid sequence shown in SEQ ID NO: 33, the LCDR2 may include the amino acid sequence shown in SEQ ID NO: 35, and the LCDR3 may include the amino acid sequence shown in SEQ ID NO: 37. For example, the reference antibody may include antibody 47A1, JYB1931A13, or an antigen-binding protein having the same LCDR3 (for example, the same LCDR1-3).

[0298] In the present application, the reference antibody may include a light chain variable region, and the light chain variable region may include the amino acid sequence shown in SEQ ID NO: 76.

[0299] DIX 3 MTQSX 8 X 9 X 10 X 11 SX 13 SVGDRVX 20 X 21 TCKASQNVGSNVAWFQQKPGX 42 X 43 PKPLIYSASYRYSGVPX 60 RFX63 GSGSGTDFTLTISX 77 X 78 QX 80 EDX 83 AX 85 YFCHQYNTYPLTFGX 100 GTKX 104 EX 106 X 107 (SEQ ID NO: 76)(where X 3 may be Q or V, X 8 may be P or Q, X 9 may be K or S, X 10 may be F or S, X 11 may be L or M, X 13 may be A or T, X 20 may be S or T, X 21 may be I or V, X 42 may be K or Q, X 43 may be A or S, X 60 may be D or S, X 63 may be S or T, X 77 may be N or S, X 78 may be L or V, X 80 may be P or S, X 83 may be F or L, X 85 may be E or T, X 100 may be A or G, X 104 may be L or V, X 106 may be I or L, X 107 may be K or N)

[0300] In the present application, the light chain variable region of the reference antibody may contain the amino acid sequence represented by any of SEQ ID NO: 31 and 47.

[0301] In the present application, the reference antibody may include HCDR1-3 and LCDR1-3. For example, the HCDR1 may include the amino acid sequence represented by SEQ ID NO: 41, the HCDR2 may include the amino acid sequence represented by SEQ ID NO: 43, the HCDR3 may include the amino acid sequence represented by SEQ ID NO: 45, the LCDR1 may include the amino acid sequence represented by SEQ ID NO: 33, the LCDR2 may include the amino acid sequence represented by SEQ ID NO: 35, and the LCDR3 may include the amino acid sequence represented by SEQ ID NO: 37. For example, the reference antibody may include antibody 47A1, JYB1931A13, or an antigen-binding protein having the same HCDR3 (for example, the same HCDR1-3) and LCDR3 (for example, the same LCDR1-3) as these.

[0302] In the present application, the reference antibody may include a heavy chain variable region and a light chain variable region. For example, the heavy chain variable region of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 13. For example, the reference antibody may include antibody 50A6 or an antigen-binding protein having the same heavy chain variable region as this. For example, the light chain variable region of the reference antibody may include the amino acid sequence represented by SEQ ID NO: 5. For example, the reference antibody may include antibody 50A6 or an antigen-binding protein having the same light chain variable region as this. For example, the reference antibody may include antibody 50A6 or an antigen-binding protein having the same heavy chain variable region and light chain variable region as this.

[0303] In the present application, the reference antibody may include a heavy chain variable region and a light chain variable region. For example, the heavy chain variable region of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 26. For example, the reference antibody may include antibody JYB1931A63 or an antigen-binding protein having the same heavy chain variable region as this. For example, the light chain variable region of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 21. For example, the reference antibody may include antibody JYB1931A63 or an antigen-binding protein having the same light chain variable region as this. For example, 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 this.

[0304] In the present application, the reference antibody may include a heavy chain variable region and a light chain variable region. For example, the heavy chain variable region of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 39. For example, the reference antibody may include antibody 47A1 or an antigen-binding protein having the same heavy chain variable region as this. For example, the light chain variable region of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 31. For example, the reference antibody may include antibody 47A1 or an antigen-binding protein having the same light chain variable region as this. For example, the reference antibody may include antibody 47A1 or an antigen-binding protein having the same heavy chain variable region and light chain variable region as this.

[0305] In the present application, the reference antibody may include a heavy chain variable region and a light chain variable region. For example, the heavy chain variable region of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 52. For example, the reference antibody may include antibody JYB1931A13 or an antigen-binding protein having the same heavy chain variable region as it. For example, the light chain variable region of the reference antibody may include the amino acid sequence shown in SEQ ID NO: 47. For example, the reference antibody may include antibody JYB1931A13 or an antigen-binding protein having the same light chain variable region as it. For example, 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. Polypeptide and immunoconjugate

[0306] In another aspect, the present application also provides one or more polypeptides that may include the isolated antigen-binding protein of the present application. For example, the polypeptide may include a fusion protein. For example, the polypeptide may include a multispecific antibody (e.g., a bispecific antibody).

[0307] In another aspect, the present application provides one or more immunoconjugates that may include the isolated antigen-binding protein of the present application. In some embodiments, the immunoconjugate may further include a pharmaceutically acceptable therapeutic agent, a marker, and / or a detection agent. Nucleic acid, vector and cell

[0308] In another aspect, the present application also provides one or more isolated nucleic acid molecules that can encode the isolated antigen-binding protein according to the present application. For example, each of the one or more nucleic acid molecules may encode the complete antigen-binding protein, or a part thereof (e.g., HCDR1-3, one or more of the heavy chain variable regions).

[0309] For example, when the nucleic acid molecules each encode a part of the antigen-binding protein, the products encoded by the nucleic acid molecules may bind to form the isolated antigen-binding protein of the present application that has functionality (e.g., can bind to MASP-2).

[0310] The nucleic acid molecule according to the present application may be isolated. For example, it can be produced or synthesized by (i) in vitro amplification such as polymerase chain reaction (PCR) amplification, (ii) cloning recombination, (iii) purification such as enzymatic cleavage and fractionation by gel electrophoresis, or (iv) synthesis such as chemical synthesis. For example, the isolated nucleic acid may be a nucleic acid molecule produced by recombinant DNA technology.

[0311] In the present application, the nucleic acid encoding the isolated antigen-binding protein can be produced by various methods known to those skilled in the art, including but not limited to obtaining the nucleic acid molecule of the isolated antigen-binding protein according to the present application using reverse transcription PCR and PCR.

[0312] In another aspect, the present application provides one or more vectors comprising one or more nucleic acid molecules according to the present application. Each vector may contain one or more of the above nucleic acid molecules. Further, the vector may also contain other genes, for example, a marker gene that enables selection of the vector in a suitable host cell and under suitable conditions. Further, the vector may further contain an expression control element that enables accurate expression of the coding region in a suitable host. Such control elements are well known to those skilled in the art and may include, for example, a promoter, a ribosome binding site, an enhancer, and other control elements that regulate gene transcription or mRNA translation. In some embodiments, the expression control sequence is an adjustable element. The specific structure of the expression control sequence may vary depending on the function of the species or cell type, but usually includes a 5' non-transcribed sequence involved in the initiation of transcription and translation, respectively, and 5' and 3' non-translated sequences such as a TATA box, a cap sequence, and a CAAT sequence. For example, the 5' non-transcribed expression control sequence may include a promoter region that may contain a promoter sequence for controlling the transcription of a functionally linked nucleic acid. The expression control sequence may include an enhancer sequence or an upstream activator sequence. In the present application, suitable promoters may include, for example, promoters for SP6, T3, and T7 polymerases, the human U6 RNA promoter, the CMV promoter, and their artificial hybrid promoters (e.g., CMV), where a portion of the promoter may be fused with a portion of the promoter of another cellular protein (e.g., human GAPDH, glyceraldehyde-3-phosphate dehydrogenase) gene and may or may not contain additional introns. One or more nucleic acid molecules according to the present application may be operably linked to the expression control element.

[0313] The vector may include, for example, a plasmid, cosmid, virus, phage, or other vectors commonly used in genetic engineering. The vector may be, for example, an expression vector. For example, the vector may be a viral vector. The viral vector may be administered directly (in vivo) to the patient, or in an indirect form, for example, cells are treated with the virus in vitro and then the treated cells are administered to the patient (ex vivo). Viral vector technology is known in the art and is described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York) and other virology and molecular biology manuals. Conventional virus-based systems may include retroviral vectors, lentiviral vectors, adenoviral vectors, adeno-associated viral vectors, and herpes simplex viral vectors for gene transfer. In some cases, gene transfer can be integrated into the host genome using retrovirus, lentivirus, and adeno-associated virus methods, and the inserted gene can be expressed long-term. Lentiviral vectors are retroviral vectors capable of introduction or infection into non-dividing cells and typically produce higher viral titers. Lentiviral vectors may include long terminal repeat 5'LTR and truncated 3'LTR, RRE, rev response element (cPPT), central terminal sequence (CTS), and / or post-translational regulatory element (WPRE). The vector according to the present application may be introduced into cells.

[0314] In another aspect, the present application provides cells. The cells may contain the isolated antigen-binding protein according to the present application, the polypeptide according to the present application, the immunoconjugate according to the present application, one or more nucleic acid molecules, and / or one or more vectors according to the present application. For example, various cells or each cell may contain one or one type of the nucleic acid molecule or vector according to the present application. For example, various cells or each cell may contain a plurality (e.g., two or more) or a plurality of types (e.g., two or more types) of the nucleic acid molecules or vectors according to the present application. For example, the vector according to the present application may be introduced into a host cell such as a prokaryotic cell (e.g., a bacterial cell), a CHO cell, an NS / 0 cell, a HEK293T cell, a 293F cell, or a HEK293A cell, or other eukaryotic cells such as plant-derived cells, fungal cells, or yeast cells. The vector according to the present application may be introduced into the host cell by a method known to those skilled in the art, such as electroporation, lipofectine transfection, lipofectamin transfection, etc. For example, the cells may contain yeast cells. For example, the cells may contain Escherichia coli cells. For example, the cells may contain mammalian cells. For example, the cells may contain immune cells.

[0315] The cells may contain immune cells. In some cases, the cells may contain immune cells. For example, the cells may contain T cells, B cells, natural killer (NK) cells, macrophages, NKT cells, monocytes, dendritic cells, granulocytes, lymphocytes, white blood cells, and / or peripheral blood monocytes. Pharmaceutical composition

[0316] In another aspect, the present application provides a pharmaceutical composition. The pharmaceutical composition may include the isolated antigen-binding protein according to the present application, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, the cell, and / or a pharmaceutically acceptable adjuvant and / or excipient. In the present application, the pharmaceutically acceptable adjuvant may include a buffer, an antioxidant, a preservative, a low molecular weight polypeptide, a protein, a hydrophilic polymer, an amino acid, a sugar, a chelating agent, a counter ion, a metal complex, and / or a nonionic surfactant. Except when not compatible with the cells according to the present application, conventional media or reagents can be considered for use in the pharmaceutical composition of the present application. In the present application, the pharmaceutically acceptable excipient may include additives other than the main drug in a pharmaceutical preparation, and is also called an auxiliary substance. For example, the excipient may include a binder, a filler, a disintegrant, and a lubricant in a tablet. As the excipient, for example, wine, vinegar, juice, etc. in a Chinese patent medicine pill may be included. The excipient may include, for example, the matrix portion of an ointment or a cream as a semi-solid preparation. The excipient may include, for example, a preservative, an antioxidant, a flavoring agent, a fragrance, a solubilizing agent, an emulsifier, a solubilizer, an osmotic pressure regulator, and a coloring agent in a liquid dosage form. Kit, use and method

[0317] In another aspect, the present application provides a method for detecting or measuring MASP-2, and the method may include using the isolated antigen-binding protein or the polypeptide.

[0318] In the present application, the method may include an in vitro method, an ex vivo method, and a method for non-diagnostic or non-therapeutic purposes.

[0319] For example, the method may include a method for detecting the presence and / or content of MASP-2 for non-diagnostic purposes, and this method includes step 1) contacting a sample with the antigen-binding protein of the present application, and Step 2) of detecting the presence and / or content of the antigen-binding protein to which the sample binds to determine the presence and / or expression level of MASP-2 in the sample from the subject may be included. In another aspect, the present application provides a kit for MASP-2, which may include using the isolated antigen-binding protein or the polypeptide.

[0320] In the present application, the kit may include an instruction manual, and the instruction manual describes a method for detecting the presence and / or content of MASP-2. For example, the method may include in vitro methods, ex vivo methods, methods for non-diagnostic or non-therapeutic purposes.

[0321] In another aspect, the present application provides the use of the isolated antigen-binding protein or the polypeptide in the manufacture of a kit that can be used in a method for detecting the presence and / or content of MASP-2. For example, the method may include in vitro methods, ex vivo methods, methods for non-diagnostic or non-therapeutic purposes.

[0322] In another aspect, the present application provides the use of the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, and the pharmaceutical composition in the prevention, remission, and / or treatment of diseases or disorders.

[0323] In another aspect, the present application provides the use of the isolated antigen-binding protein, the polypeptide, the immunoconjugate, the isolated nucleic acid molecule, the vector, the cell, and / or the pharmaceutical composition in the manufacture of a drug for preventing, remitting, and / or treating a disease or disorder.

[0324] In another aspect, the present application provides a method for preventing and / or treating a disease or disorder, including administering the isolated antigen-binding protein, the isolated nucleic acid molecule, the vector, the cell, and the pharmaceutical composition to a subject in need thereof.

[0325] The pharmaceutical compositions, pharmaceutical combinations, and methods of the present application can be used in combination with other types of cancer therapies such as chemotherapy, surgery, radiotherapy, gene therapy, etc. The pharmaceutical compositions and methods described in the present application are useful for other disease states that depend on immune responses such as inflammation, immune diseases, and infectious diseases.

[0326] In the present application, the subject may include a human or a non-human animal. The non-human animal may be selected from the group consisting of, for example, monkeys, chickens, geese, cats, dogs, mice, and rats. Also, the non-human animal can include animal species other than humans, such as livestock animals, rodents, primates, captive animals, poultry, etc. The human may be white, African, Asian, Semitic, other ethnic groups, or a mixture of ethnic groups. Further, for example, the human may be an elderly person, an adult, a teenager, a child, or an infant.

[0327] The effective dose in humans can be estimated from the effective dose in experimental animals. For example, Freireich et al. have described the dose correlation between animals and humans (based on milligrams per square meter of body surface area) (Freireich et al., Cancer Chemother. Rep. 50, 219 (1966)). The body surface area can be approximately determined from the height and weight of the patient. See, for example, Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y., 537 (1970).

[0328] Without being limited to any theory, the following examples are used only to illustrate the fusion proteins of the present application, their production methods, uses, etc., and are not used to limit the scope of the invention of the present application. Examples Example 1 Production of Antigen

[0329] Human MASP-2 protein (O00187) contains six functional domains. In this example, functional domain 4 (CPP1), functional domain 5 (CPP2), and functional domain 6 (SP) were selected and recombinantly expressed in E. coli. Since the expressed protein existed as inclusion bodies, the inclusion bodies were refolded to obtain the active protein, and then purified by the His at the C-terminus of the protein fragment. The obtained recombinant human MASP-2 proteins were hMASP2-D456 (amino acid sequence shown in SEQ ID NO: 1) and hMASP2A-D456 (amino acid sequence shown in SEQ ID NO: 2), respectively. Using the same production method, MASP-2 of cynomolgus monkey (A0A2K5UJY0, cMASP2-D456 having the amino acid sequence shown in SEQ ID NO: 4) and mouse (Q91WP0, mMASP2-D456 having the amino acid sequence shown in SEQ ID NO: 3) was produced. Example 2 Production of Mouse Anti-Human MASP-2 Monoclonal Antibody

[0330] 2.1 Animal Immunization At the first immunization, using complete Freund's adjuvant, hMASP2A-D456 and PADRE were emulsified at a ratio of 1:1, and then intraperitoneally injected into 6- to 8-week-old female SD rats at 50 μg hMASP2A-D456 per rat, and injected into multiple sites in the peritoneal cavity and subcutaneous of 6- to 8-week-old female Balb / c mice. Thereafter, booster immunizations were performed at intervals of 2 to 3 weeks. Each rat was intraperitoneally injected with 50 μg of the antigen and incomplete Freund's adjuvant, and injected into multiple sites in the peritoneal cavity and subcutaneous of the mice. Each mouse was intraperitoneally injected with 50 μg of the antigen and incomplete Freund's adjuvant. A total of 3 immunizations were performed. Two weeks after the last immunization, blood was collected from the tails of the animals, and the anti-human MASP-2 antibody titer of the serum was measured. After performing shock immunization by intraperitoneally injecting 50 μg of the protein without Freund's adjuvant into the animals, spleen cells were excised on the 3rd day. 2.2 Spleen Cell Fusion After euthanizing the rats / mice, dissection, spleen extraction, and pulverization were performed to collect cells, which were centrifuged at 1500 rpm for 5 minutes to collect the cells. The cells were suspended in 5 - 10 milliliters of erythrocyte lysate, left at 4°C for 10 minutes, and the reaction was stopped with DMEM + 10% FBS and counted. After centrifugation, the cells were suspended in 40 ml of DMEM, allowed to stand for 2 - 3 minutes, and then the supernatant was transferred to another 50 ml centrifuge tube. SP2 / 0 cells were collected and mixed with spleen cells at a ratio of SP2 / 0: spleen cells = 1:2, centrifuged, and after thoroughly aspirating the supernatant, pipetting was performed to precipitate the mixed cells. The mixed cells were washed twice with DMEM, and PEG fusion was carried out by the conventional method. After fusion, the cells were washed with DMEM medium and resuspended in a screening medium of DMEM + 10% FBS + 1×HAT. The fused cells were added to a 96-well cell culture plate and incubated in an incubator at 37°C, 75% humidity, and 5% CO 2 and cultured for 9 - 10 days. 2.3 Screening of mouse monoclonal antibodies (1) Identification of the binding function of the supernatant of hybridoma monoculture to hMASP2-D456 by ELISA hMASP2-D456 was diluted to 1.0 μg / mL with carbonate buffer and added to a high-binding clear polystyrene 96-well plate (Nunc) at 100 μL / well, followed by overnight coating at 4 °C. The next day, the ELISA plate was washed twice with washing buffer (PBS + 0.05% Tween 20 (sigma)) using an automatic washer. 300 μL of blocking buffer (PBS + 0.05% Tween 20 (sigma) + 1% BSA) was added per well and blocked at room temperature for 1 hour. Then, it was washed twice with washing buffer using an automatic washer, 1000 μL of hybridoma supernatant was transferred to each well of the ELISA plate, incubated at room temperature for 1 hour, and then the plate was washed 3 times by the above method. Per well, 100 μL of goat anti-mouse HRP (Sigma, product number: M4280) and rabbit anti-rat Fc-HRP (Sigma, product number: A5795) diluted 1:5000 with blocking buffer were added. After incubation at room temperature for 1 hour, the plate was washed 3 times by the above method. 100 μL / well of TMB substrate solution was added, and then 50 μL of 1.0 M hydrochloric acid stop solution was added per well to stop the reaction, and it was read at 450 nm using a Thermo Multiscan FC. By the binding experiment, 50A6 was screened as a hybridoma cloning rat and 47A1 was screened as a hybridoma cloning mouse. Using serum-free culture and the conventional antibody purification method, a hybridoma monoclonal antibody was prepared and the antibody function was confirmed. Example 3 Identification of Mouse Anti-Human MASP-2 Monoclonal Antibody

[0331] 3.1 Identification of the Binding Function of Mouse Anti-Human MASP-2 Monoclonal Antibody to Human MASP-2 by ELISA hMASP2-D456 was diluted to 1.0 μg / mL and 0.5 μg / mL with carbonate buffer solution (CBS), added to a high-binding clear polystyrene 96-well plate (Nunc) at 100 μL / well, and coated overnight at 4°C. The next day, the ELISA plate was washed twice with washing buffer (PBS + 0.05% Tween 20 (sigma)) using an automatic washer. 300 μL of blocking buffer (PBS + 0.05% Tween 20 (sigma) + 1% BSA) was added per well and blocked at room temperature for 1 hour. Then, it was washed twice with washing buffer using an automatic washer, and the monoclonal antibody was serially diluted 8-fold with blocking buffer. It was sequentially added to each well of the ELISA plate, incubated at room temperature for 1 hour, and then the plate was washed 3 times by the above method. 100 μL of goat anti-mouse HRP (Sigma, product number: M4280) and rabbit anti-rat Fc-HRP (Sigma, product number: A5795) diluted 1:5000 with blocking buffer was added per well. After incubation at room temperature for 1 hour, the plate was washed 3 times by the above method. 100 μL of TMB substrate solution was added per well, and then 50 μL of 1.0 M hydrochloric acid stop solution was added per well to stop the reaction, and it was read at 450 nm on a Thermo Multiscan FC. It was plotted using Graphpad and the EC50 was calculated. As a result, as shown in Figure 1, the 50A6 antibody was able to bind to human MASP-2. As shown in Figure 2, the 47A1 antibody was able to bind to human MASP-2, and all of the mouse anti-human MASP-2 monoclonal antibodies of the present application were able to bind to human MASP-2. 3.2 Identification of the cross-binding function of mouse anti-human MASP-2 monoclonal antibodies to mouse mMASP2-D456 and monkey cMASP2-D456 by ELISA Mouse mMASP2-D456 and cynomolgus monkey cMASP2-D456 were diluted to 1.0 μg / mL and 0.5 μg / mL with carbonate buffer, and added to a high-binding clear polystyrene 96-well plate (Nunc) at 100 μL / well, and coated overnight at 4°C. The next day, the ELISA plate was washed twice with washing buffer (PBS + 0.05% Tween 20 (sigma)) using an automatic washer. 300 μL of blocking buffer (PBS + 0.05% Tween 20 (sigma) + 1% BSA) was added per well and blocked at room temperature for 1 hour. Then, it was washed twice with washing buffer using an automatic washer, and the monoclonal antibody 50A6 was diluted 11-fold to 15.0 μg / mL with blocking buffer. It was sequentially added to each well of the ELISA plate, incubated at room temperature for 1 hour, and then the plate was washed 3 times by the above method. Per well, 100 μL of goat anti-mouse HRP (Sigma, product number: M4280) and rabbit anti-rat Fc-HRP (Sigma, product number: A5795) diluted 1:5000 with blocking buffer were added. After incubation at room temperature for 1 hour, the plate was washed 3 times by the above method. 100 μL of TMB substrate solution was added per well, and then 50 μL of 1.0 M hydrochloric acid stop solution was added per well to stop the reaction. It was read at 450 nm using a Thermo Multiscan FC. It was plotted using Graphpad and the EC50 was calculated. The results of the binding of 50A6 to cynomolgus monkey cMASP2-D456 are shown in Figure 3A, and the results of the binding of 50A6 to mouse mMASP2-D456 are shown in Figure 3B. The results of the binding of 47A1 to cynomolgus monkey cMASP2-D456 are shown in Figure 4A, and the results of the binding of 47A1 to mouse mMASP2-D456 are shown in Figure 4B. By identifying functions such as binding and blocking, 50A6 was humanized as a rat-derived candidate antibody, and 47A1 was humanized as a mouse-derived candidate antibody. Example 4 Humanization of Hybridoma Monoclonal Antibodies

[0332] 4.1 The human germline sequence (data source: IMGT) most homologous to the mouse-derived antibody 50A6 was used as the humanized design framework (the light chain used GKV6-21*02 and IGKJ2*01 as the framework, and the heavy chain used IGHV3-7*01 and IGH4*01 as the framework) through sequence alignment. For the mouse-derived antibody 47A1, IGKV1-16*01, IGKJ4*01, and IGHV4-30-4*01, IGHJ4*01 were used as the frameworks for the light and heavy chains, respectively. The variable regions of the light and heavy chains of the antibody were numbered according to Chothia (see Chothia & Lesk, 1987), and the CDR regions of the antibody were defined as follows: CDRL1 (L24-L34, i.e., amino acids 24 to 34 of VL), CDRL2 (L50-L56), CDRL3 (L89-L97), CDRH1 (H26-H32, i.e., amino acids 26 to 32 of VH), CDRH2 (H52-H56), CDRH3 (H95-H97). Based on sequence alignment and the structural information of the variable regions, humanized mutations were introduced into the amino acids of the variable regions of the light and heavy chains of the antibody. 4.2 An expression vector was designed, the gene was synthesized, and recombinant antibodies were expressed and purified in mammalian cells. The differences in antibody activity and physicochemical properties after humanization modification were compared, and humanization optimization was performed 1 to 2 times. 4.3 CDR transplantation was performed using the above germline antibody as the framework. First, the variable region amino acid sequences of the following chimeric antibodies were obtained: the light chain variable region sequence of 50A6 (the amino acid sequence shown in SEQ ID NO: 5), the heavy chain variable region sequence of 50A6 (the amino acid sequence shown in SEQ ID NO: 13). The light chain variable region sequence of 47A1 (the amino acid sequence shown in SEQ ID NO: 31), the heavy chain variable region sequence of 47A1 (the amino acid sequence shown in SEQ ID NO: 39). Furthermore, amino acid mutations were introduced based on the variable region amino acid sequences of the above chimeric antibodies to obtain the humanized optimized amino acid sequences of the light chain variable region sequence of JYB1931A63 (the amino acid sequence shown in SEQ ID NO: 21) and the heavy chain variable region sequence of JYB1931A63 (the amino acid sequence shown in SEQ ID NO: 26). The light chain variable region sequence of JYB1931A13 (amino acid sequence shown in SEQ ID NO: 47), the heavy chain variable region sequence of JYB1931A13 (amino acid sequence shown in SEQ ID NO: 52). 4.4 Expression and purification of antibodies After designing the above humanized optimized amino acid sequences, codon optimization was performed respectively, and the corresponding DNA gene fragments (Genscript) were synthesized. The synthesized gene fragments were amplified into the expression vector pcDNA3.4 (Life Technologies). After amplification of the expression plasmid and plasmid extraction, the double plasmid was co-transfected into ExpiCHO cells (ThermoFisher Scientific, A29133), and transient expression of the antibody was performed as follows according to the method of the supplier ExpiCHO expression system. That is, in a total culture volume of 25 mL of medium, ExpiCHO cells were cultured at 36.5 °C and a carbon dioxide concentration of 8% until the density reached 6×10 6 / mL. Using the ExpiFectamine transfection reagent, 10 μg each of the light and heavy chain expression plasmids of the antibody were transfected into the cells. One day after transfection, 150 μL each was taken and added to the cultured cells together with 4 mL of ExpiCHO enhancer and ExpiCHO adjuvant, and the culture was continued for up to 9 days. Then, centrifugation was performed at 4 °C and 3500 rpm, and the supernatant was collected. AmMagTM Protein A beads (Genscript, L00695) were mixed with the antibody expression supernatant, incubated at room temperature for 2 hours, washed twice with PBS, the supernatant was discarded, an appropriate amount of elution buffer Protein G or A SefinoseTM Elution buffer (Sangon, C600481) was added, and after thorough mixing, it was placed in a test tube rack and allowed to stand for 5 min incubation. During the incubation period, the magnetic beads were resuspended 2 - 3 times, elution was repeated twice. Immediately after elution, an appropriate amount of neutralizing solution 1M Tris-HCl, pH 7.5 (Sangon, B548124) was added for neutralization. Example 5 Blocking of the lectin pathway of the human complement system by an anti-MASP-2 mouse-derived antibody

[0333] 5.1 Detection of the blockage of the human serum complement signaling pathway by antibodies was performed using the WIESLAB (registered trademark) complement system lectin pathway (Svar Life Science AB, product number: AS 1327) kit. Human serum and antibodies 50A6 and 47A1 were diluted with three types of diluents. Here, the dilution methods for human serum were CP: 80 ul / 4 ml, MP: 80 ul / 4 ml, and AP: 444.7 / 4 ml, and the antibodies were serially diluted 10-fold from 500 nM. 5.2 Experimental steps First, 50 μL of each of the diluted samples and serum were added to the plate and incubated at 37 °C for 1 hour. 2) The plate was washed three times with the wash buffer. 3) 100 μL of coupling was added per well and left at room temperature for 30 min. 4) The plate was washed, i.e., the plate was washed three times with the wash buffer. 5) 100 μL of the substrate solution was added per well and left at room temperature for 30 min. 6) 100 μL of 5 mM EDTA was added per well to stop the color development. 7) After stopping the color development, the absorbance at 450 nm was read using a multimode microplate reader. 5.3 Data processing: The data were processed using GraphPad Prism software. As shown in Figure 5, the antibody of the present application can block the human complement system lectin pathway. Example 6 Measurement of the affinity of the anti-MASP-2 antibody

[0334] 6.1 Sample dilution The affinities of the candidate antibodies JYB1931A63 and JYB1931A13 for human hMASP2-D456 (KACTUS, lot number: 080203), cynomolgus monkey cMASP2-D456 (KACTUS, lot number: 030301), and mouse mMASP2-D456 (KACTUS, lot number: 030401) were measured using Octet RED96e (manufactured by Fortebio). Both the antigen and the antibody were diluted with 1×PBST (1×PBS: Sangon Biotech, B548117-0500; 0.02% Tween 20: sigma-aldrich, P1379), and the concentration of the antigen was 30 nM and the concentration of the antibody was 5 μg / mL. 6.2 On-machine Detection of Samples (Octet Data Acquisition 11.1.0.11) First, the sample was added to a 96-well plate (Greiner bio-one, 655209) at 200 μL / well. Next, the software parameters were set, the plate temperature was set to 30 °C, and the frequency for collecting the standard kinetic signal was set to 5.0 Hz. Next, the AHC sensor (manufactured by Fortebio, product number: 18-0015) was pre-wetted with 1×PBST for 10 min and then detected by the machine. Each cycle included the following steps. 1) Immerse in the buffer for 60 s. 2) Detect whether the antigen is non-specifically bound to the sensor. 3) Regenerate with a 10 mM glycine solution at pH 1.7. 4) Immerse in the buffer for 60 s. 5) Immobilize the antibody on the sensor in 60 s. 6) Immerse the sensor in the buffer for 180 s. 7) Bind the antigen and the antibody for 180 s. 8) Dissociate the antigen and the antibody for 5 min. 9) Regenerate the sensor. 6.3 Data Analysis Using the Data Analysis 12.0 software manufactured by Fortebio, for the form in which the antigen-antibody binds in a 1:1 ratio, by measuring the association rate (Ka) and the dissociation rate (Kd), the equilibrium dissociation constant (K D ) of the antibody was calculated. The results are shown in Table 1.

Table 1

[0335] Using the WIESLAB (registered trademark) complement system lectin pathway (Svar Life Science AB, product number: AS 1327) kit, the blocking of the complement signaling pathway of human serum and monkey serum by antibodies was detected. Human serum and monkey serum were diluted with three types of diluents. Here, the dilution means for human serum were CP: 80ul / 4ml, MP: 80ul / 4ml, and AP: 444.7 / 4ml. The dilution means for monkey serum were CP: 2 / 100, MP: 2 / 100, and AP: 11.1 / 100. For each sample, it was serially diluted 10-fold from 500 nM with three types of diluents. The experimental steps and data processing were the same as in Example 5. As shown in Figure 6, the humanized antibody of the present application was able to block the complement system lectin pathway. Example 8 Detection of Physicochemical Properties of Antibodies

[0336] 8.1 SEC-HPLC Purity Analysis (1) The sample was diluted to 1 mg / mL, mixed uniformly, centrifuged at 12,000 rpm for 5 min, the supernatant was taken and transferred to a vial, and placed in an HPLC sample dish. The chromatographic conditions were set as shown in Table 2. [Table 2] (2) After equilibrating the column with the mobile phase (200 mM phosphate buffer, pH 6.8), the sample was injected for analysis, data analysis was performed with chromatographic software, and the peak area percentage of each peak was calculated by the peak area normalization method. 8.2 HIC-HPLC Analysis (1) The sample was diluted to 1 mg / ml, centrifuged, and the supernatant was collected for measurement. The chromatographic conditions were set as follows (Table 3). [Table 3] (2) Gradient elution was performed with mobile phase A (50 mM phosphate buffer / 1 M ammonium sulfate, pH 7.0) and mobile phase B (50 mM phosphate buffer, pH 7.0), and the main peak retention time was recorded. 8.3 Analysis of Melting Temperature (Tm) Value After diluting the test article to 1 mg / mL with sample buffer, Protein Thermal Shift TM According to the handling instructions of the Protein Thermal Shift Starter Kit, add 13 μL of the test article solution to a PCR tube, add 5 μL of Protein Thermal shift buffer, add 2 μL of 10× staining solution to make the reaction volume 20 μL, mix uniformly, and then centrifuge at 12,000 rpm for 5 min to remove air bubbles. Place the detection sample in a PCR instrument, perform sample analysis, and record the Tm value of the sample. TM 8.4 iCIEF analysis Add the sample solution to a system obtained by thoroughly mixing 70 μl of 1% methylcellulose (MC), 80 μl of 5 M urea, 8 μl of amphoteric electrolyte Pharmalyte pH 3 - 10, 1 μl each of pI marker 5.5 and pI marker 9.5. Supplement with an appropriate amount of ultrapure water to 200 μl, and mix uniformly. Centrifuge and collect the supernatant, inject and analyze. After the analysis is completed, import the result file into ChromPerfect software for spectral integration processing, and calculate the isoelectric point of each peak and the percentage of each peak, which are shown in Table 4.

Table 4

[0337] Humanized FcRn mice were used as experimental animals, and the pharmacokinetic indices after single intraperitoneal administration of two test drug positive controls (Narsoplimab) and JYB1931A63 were studied respectively. All animal experiment programs were reviewed and approved by IACUC. The hFcRn mice were purchased from Beijing Biocytogen Co., Ltd., male, 6 - 8 weeks old, weighing 23 - 26 g, housed in an SPF - grade animal room, fed with standard pellet feed, allowed to freely ingest food and water, at room temperature of 18 - 24 °C, relative humidity of 40% - 50%, and the day and night were alternated for 12 hours each day. There were 16 experimental animals in total, randomly divided into 4 groups of 4 animals each, and administered intraperitoneally once. The dosage was 10 mg / kg, and the administration volume was 10 mL / kg. The blood sampling time points were before administration, 2 h, 6 h, 24 h (day 1), day 2, day 3, day 4, day 7, day 10, day 14, day 21, day 28, day 35, and day 42 after administration. 60 μL of whole blood per animal was collected from the orbital cavity into an EP tube, left standing at room temperature for 30 min, and then centrifuged (2000 g, 4 °C, 5 min) to separate the serum. Each sample was divided into two parts (detection tube and backup tube), 10 μL per tube, and stored at - 80 °C. The indirect Elisa method was used to analyze the pharmacokinetics of the four drugs at each time point. Recombinant human MASP - 2 protein was used as the antigen, coated at 1 μg / mL, 100 μL / well, and left at 4 °C overnight. After washing the plate, it was blocked at 4 °C overnight with 200 μL / well of blocking solution. Serum samples were added at 50 μL / well, incubated at 37 °C for 1 h. The detection antibody was mouse monoclonal antibody [H2] anti - human IgG F(ab)’2 (HRP) (abcam, ab87422, GR3246767 - 11)+Streptavidin - peroxidase (Sigma, lot: SLCB5784), 100 μl / well, incubated at 37 °C for 0.5 h. Color development was carried out with TMB chromogenic solution (KPL, product number: 52 - 00 - 03), and the OD450 value was read with a microplate reader (Molecular Devices, SpectraMax M3). The drug concentration was obtained from the standard curve, and the PK parameters were obtained using non - compartmental analysis of PK Solver. The blood concentration curve is shown in Figure 7.

[0338] When the positive control was administered, the half-life (t 1 / 2 ) = 4.93 days, the peak time point (Tmax) was 0.45 days, and the maximum concentration (Cmax) was 9393 ng / ml. When JYB1931A63 was administered, the half-life (t 1 / 2 ) = 18.6 days, the peak time point (Tmax) was 4.53 days, and the maximum concentration (Cmax) was 86934 ng / ml.

[0339] The foregoing detailed description is provided by way of explanation and illustration and is not intended to limit the appended claims. Various changes to the embodiments set forth in this application will be apparent to those skilled in the art and are intended to be included within the scope of the appended claims and their equivalents.

Claims

**Claim 1**: An isolated antigen-binding protein that specifically binds to cynomolgus macaque MASP-2 protein with a KD value of about 2E-09 M or less in Octet detection, comprising a heavy chain variable region VH and a light chain variable region VL, wherein the heavy chain variable region VH comprises HCDR1, HCDR2, and HCDR3, of which the HCDR3 comprises the amino acid sequence represented by SEQ ID NO: 19, the HCDR2 comprises the amino acid sequence represented by SEQ ID NO: 17, the HCDR1 comprises the amino acid sequence represented by SEQ ID NO: 15, the light chain variable region VL comprises LCDR1, LCDR2, and LCDR3, of which the LCDR3 comprises the amino acid sequence represented by SEQ ID NO: 11, the LCDR2 comprises the amino acid sequence represented by SEQ ID NO: 9, and the LCDR1 comprises the amino acid sequence represented by SEQ ID NO: 7, an isolated antigen-binding protein. **Claim 2**: Comprising H-FR1, H-FR2, H-FR3, and H-FR4, of which the H-FR1 comprises the amino acid sequence represented by SEQ ID NO: 62, the H-FR2 comprises the amino acid sequence represented by SEQ ID NO: 63, the H-FR3 comprises the amino acid sequence represented by SEQ ID NO: 64, and the H-FR4 comprises the amino acid sequence represented by SEQ ID NO: 65, or, the H-FR1, H-FR2, H-FR3, and H-FR4 comprise any one set of amino acid sequences selected from the following a to b, the isolated antigen-binding protein according to claim 1. a) H-FR1: SEQ ID NO: 27, H-FR2: SEQ ID NO: 28, H-FR3: SEQ ID NO: 29, and H-FR4: SEQ ID NO: 30, b) H-FR1: SEQ ID NO: 14, H-FR2: SEQ ID NO: 16, H-FR3: SEQ ID NO: 18, and H-FR4: SEQ ID NO:

20. **Claim 3**: Comprising a heavy chain variable region VH comprising the amino acid sequence represented by SEQ ID NO: 67, or, the VH comprises the amino acid sequence represented by any of SEQ ID NO: 26 and SEQ ID NO: 13, the isolated antigen-binding protein according to claim 1 or 2. **Claim 4** A protein according to claim 1, comprising L-FR1, L-FR2, L-FR3, and L-FR4, wherein L-FR1 comprises the amino acid sequence shown in SEQ ID NO: 58, L-FR2 comprises the amino acid sequence shown in SEQ ID NO: 59, L-FR3 comprises the amino acid sequence shown in SEQ ID NO: 60, and L-FR4 comprises the amino acid sequence shown in SEQ ID NO:

61. Or, the L-FR1, L-FR2, L-FR3, and L-FR4 comprise any one set of amino acid sequences selected from the following a to b, and the isolated antigen-binding protein according to claim 1. a) L-FR1: SEQ ID NO: 22, L-FR2: SEQ ID NO: 23, L-FR3: SEQ ID NO: 24, and L-FR4: SEQ ID NO: 25 b) 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. **Claim 5** The VL comprises the amino acid sequence shown in SEQ ID NO: 66, Or, the VL comprises the amino acid sequence shown in either SEQ ID NO: 21 or SEQ ID NO: 5, and the isolated antigen-binding protein according to claim 1. **Claim 6** A protein according to claim 3 or 5, comprising VH and VL, wherein the VH and VL comprise any one set of amino acid sequences selected from the following a to b. a) VH: SEQ ID NO: 26, and VL: SEQ ID NO: 21 b) VH: SEQ ID NO: 13, and VL: SEQ ID NO: 5 **Claim 7** Comprising a heavy chain constant region comprising a constant region derived from IgG or a constant region derived from IgY, Or, the heavy chain constant region comprises a constant region derived from IgG, Or, the heavy chain constant region comprises a constant region derived from IgG1, IgG2, IgG3, or IgG4, Or, the heavy chain constant region comprises the amino acid sequence shown in SEQ ID NO: 56, and the isolated antigen-binding protein according to claim 1. **Claim 8** Comprising a light chain constant region comprising a constant region derived from Igκ or a constant region derived from Igλ, Or, the light chain constant region comprises a constant region derived from human Igκ, Or, the isolated antigen-binding protein according to claim 1, wherein the light chain constant region comprises the amino acid sequence represented by SEQ ID NO:

57.

9. Comprising an antibody or an antigen-binding fragment thereof, The isolated antigen-binding protein according to claim 1, wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab’, F(ab)2, Fv fragment, F(ab’)2, scFv, and di-scFv.

10. The isolated antigen-binding protein according to claim 9, wherein the antibody is selected from the group consisting of monoclonal antibody, single-chain antibody, chimeric antibody, humanized antibody, and fully human antibody.

11. A polypeptide comprising the isolated antigen-binding protein according to any one of claims 1 to 10.

12. An immunoconjugate comprising the isolated antigen-binding protein according to any one of claims 1 to 10 or the polypeptide according to claim 11.

13. An isolated nucleic acid molecule encoding the isolated antigen-binding protein according to any one of claims 1 to 10 or the polypeptide according to claim 11.

14. A vector comprising the isolated nucleic acid molecule according to claim 13.

15. A cell comprising the isolated antigen-binding protein according to any one of claims 1 to 10, the polypeptide according to claim 11, the immunoconjugate according to claim 12, the isolated nucleic acid molecule according to claim 13, and / or the vector according to claim 14.

16. A method for producing the isolated antigen-binding protein according to any one of claims 1 to 10 or the polypeptide according to claim 11, The method comprising culturing the cell according to claim 15 under conditions for expressing the isolated antigen-binding protein according to any one of claims 1 to 10 or the polypeptide according to claim 11.

17. A pharmaceutical composition comprising the isolated antigen-binding protein according to any one of claims 1 to 10, the polypeptide according to claim 11, the immunoconjugate according to claim 12, the isolated nucleic acid molecule according to claim 13, the vector according to claim 14, the cell according to claim 15, and / or a pharmaceutically acceptable adjuvant and / or excipient.

18. A method for detecting or measuring MASP-2, comprising using the isolated antigen-binding protein according to any one of claims 1 to 10 or the polypeptide according to claim 11.

19. A detection kit for MASP-2, comprising the isolated antigen-binding protein according to any one of claims 1 to 10 or the polypeptide according to claim 11.

20. A preparation comprising the isolated antigen-binding protein according to any one of claims 1 to 10, the polypeptide according to claim 11, the immunoconjugate according to claim 12, the isolated nucleic acid molecule according to claim 13, the vector according to claim 14, the cell according to claim 15 and / or the pharmaceutical composition according to claim 17, which is used for the prevention, remission and / or treatment of diseases or disorders.

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