Antibodies targeting MASP-2 and uses thereof

Abstract

Disclosed herein are anti-MASP-2 antibodies and antigen-binding fragments, polynucleotides encoding the antibodies and antigen-binding fragments, and pharmaceutical compositions comprising the antibodies and antigen-binding fragments. Also disclosed are the use of the anti-MASP-2 antibodies and antigen-binding fragments described herein in the treatment of conditions and disorders associated with complement activation.

Description

Detailed Description of the Invention

[0001] This application claims priority from PCT / CN2022 / 081803, filed March 18, 2022, the entire contents of which are incorporated herein by reference. 1. Field

[0001] The present invention relates to molecular biology and immunology. Provided herein are anti-MASP-2 antibodies and their use in the treatment of conditions associated with complement activation. 2.Background

[0002] Mannan-binding lectin-associated serine proteinase 2 (MASP-2) is an effector enzyme required for activation of the lectin pathway of complement. In addition to its essential role in immune defense, inappropriate or uncontrolled activation of the lectin pathway contributes to tissue damage in many clinical conditions. However, there has been limited success in developing therapeutic complement inhibitors that target the lectin pathway. Thus, there is an unmet need for additional treatment options for agents that target MASP-2. The compositions and methods provided herein fulfill these needs and offer other relative advantages. 3. Overview

[0003] As used herein, an antibody or antigen-binding fragment thereof that specifically binds to human MASP-2 comprises: (1) a light chain variable region (VL) comprising, as defined by Kabat, (a) a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9, and 11, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VL CDRs; and / or (b) a heavy chain variable region (VH) comprising, as defined by IMGT, (a) a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 12, 16, and 18, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDRs; or (2) a heavy chain variable region (VH) comprising, as defined by IMGT, (a) a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 8, 10, and 11, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDRs. The present invention provides an antibody or antigen-binding fragment thereof comprising: (a) a VL comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 13, 17, and 19, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDRs; and / or (b) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 13, 17, and 19, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDRs.

[0002]

[0004] In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a VL CDR1, a VL CDR2, a VL CDR3, a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9, 11, 12, 16, and 18, respectively, as defined by Kabat.

[0003]

[0005] In some embodiments, the antibodies or antigen-binding fragments provided herein, as defined by IMGT, comprise a VL CDR1 having the amino acid sequence of SEQ ID NO: 8, a VL CDR2 having the amino acid sequence of SEQ ID NO: 10, a VL CDR3 having the amino acid sequence of SEQ ID NO: 11, a VH CDR1 having the amino acid sequence of SEQ ID NO: 13, 14, or 15, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 19 or 20.

[0004]

[0006] In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a VL CDR1, a VL CDR2, a VL CDR3, a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of: (1) SEQ ID NOs: 8, 10, 11, 13, 17, and 19, respectively; (2) SEQ ID NOs: 8, 10, 11, 14, 17, and 19, respectively; (3) SEQ ID NOs: 8, 10, 11, 15, 17, and 19, respectively; (4) SEQ ID NOs: 8, 10, 11, 13, 17, and 20, respectively; (5) SEQ ID NOs: 8, 10, 11, 14, 17, and 20, respectively; or (6) SEQ ID NOs: 8, 10, 11, 15, 17, and 20, respectively.

[0005]

[0007] Also provided herein is an antibody or antigen-binding fragment thereof that specifically binds to human MASP-2, comprising (a) a VL having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 21, and / or (b) a VH having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 22.

[0006]

[0008] In some embodiments, the antibody or antigen-binding fragment provided herein comprises a VL having the amino acid sequence of SEQ ID NO:21 and a VH having the amino acid sequence of SEQ ID NO:22.

[0007]

[0009] The present specification also provides an antibody or antigen-binding fragment thereof that specifically binds to human MASP-2, comprising (a) a VL comprising VL CDR1, VL CDR2 and VL CDR3 derived from a VL having the amino acid sequence of SEQ ID NO: 21, and / or (b) a VH comprising VH CDR1, VH CDR2 and VH CDR3 derived from a VH having the amino acid sequence of SEQ ID NO: 22.

[0008]

[0010] In some embodiments, the antibodies or antigen-binding fragments provided herein are chimeric antibodies or antigen-binding fragments, humanized antibodies or antigen-binding fragments, or human antibodies or antigen-binding fragments.

[0009]

[0011] In some embodiments, the antibodies or antigen-binding fragments provided herein are humanized antibodies or antigen-binding fragments. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise (a) a VL having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 23, and / or (b) a VH having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 24. In some embodiments, the antibodies or antigen-binding fragments provided herein comprise a VL having the amino acid sequence of SEQ ID NO: 23 and a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 24-32.

[0010]

[0012] In some embodiments, the antibody or antigen-binding fragment provided herein is selected from the group consisting of Fab, Fab', F(ab')2, Fv, scFv, (scFv)2, single domain antibody (sdAb), and heavy chain antibody (HCAb).

[0011]

[0013] In some embodiments, the antibodies or antigen-binding fragments provided herein are IgG1 antibodies or variants thereof, IgG2 antibodies or variants thereof, IgG3 antibodies or variants thereof, or IgG4 antibodies or variants thereof.

[0012]

[0014] In some embodiments, an IgG4 antibody or variant thereof provided herein comprises (1) a light chain having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 33, and (2) a heavy chain having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 34.

[0013]

[0015] In some embodiments, the heavy chain of an antibody provided herein has an amino acid sequence selected from the group consisting of SEQ ID NOs: 34-42, or a variant thereof modified by one or more amino acid substitutions that increase the terminal half-life of the antibody. In some embodiments, the heavy chain variant is modified by one or more substitutions at amino acid residues selected from the group consisting of S228, F234, L235, M252, S254, T256, K288, T307, M428, N434, H435, and Y436 (numbered according to the EU index). In some embodiments, the heavy chain variants are modified by amino acid substitutions selected from the group consisting of: i) S228P, ii) F234A and L235A, iii) S228P, F234A and L235A, iv) T307H and N434A, v) M252Y, S254T and T256E, vi) M428L, N434A and Y436T, vii) S228P, M252Y, S254T and T256E, viii) S228P, F234A, L235A, M252Y, S254T and T256E, ix) S228P, F234A, L235A, T307Q and N434A, and (x) M252Y, S254T, T307H and N434A.

[0014]

[0016] In some embodiments of the antibodies provided herein, the light chain has the amino acid sequence of SEQ ID NO: 33 and the heavy chain has an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-74.

[0015]

[0017] Also provided herein are antibodies or antigen-binding fragments thereof that compete with the antibodies or antigen-binding fragments described herein for binding to human MASP-2.

[0018] In some embodiments, the antibodies or antigen-binding fragments provided herein are bispecific or multispecific antibodies.

[0016]

[0019] In some embodiments, the antibodies or antigen-binding fragments provided herein are monoclonal antibodies or antigen-binding fragments.

[0020] In some embodiments, the antibodies or antigen-binding fragments provided herein have: (1) a K of about 1 nM or less as measured by surface plasmon resonance (SPR); D (2) binds to human MASP-2 with an IC of approximately 0.1 μg / mL or less measured in vitro 50 (3) an IC of approximately 0.1 μg / mL or less measured in vitro; 50 (4) an IC of approximately 0.1 μg / mL or less measured in vitro; 50 (5) inhibiting MAC activation; (6) not affecting the classical or alternative pathways of complement activation; or (7) having any combination of the properties of (1) to (5).

[0017]

[0021] In some embodiments, the antibody or antigen-binding fragment provided herein specifically binds to the protease domain of human MASP-2, and the antibody or antigen-binding fragment has (1) a K of 1 nM or less as measured by SPR. D binds to human MASP-2 with an IC of 0.1 μg / mL or less measured in vitro 50 (3) an IC of 0.1 μg / mL or less measured in vitro; 50(4) an IC of 0.1 μg / mL or less measured in vitro; 50 (5) inhibiting MAC activation; (6) not affecting the classical or alternative pathways of complement activation; or (7) having any combination of the properties of (1) to (5).

[0018]

[0022] In some embodiments, the antibodies or antigen-binding fragments provided herein have an IC of 0.001 to 0.01 μg / mL measured in vitro. 50 In some embodiments, the antibodies or antigen-binding fragments provided herein have an IC of 0.1 μg / mL or less measured in vitro in the presence of 5-50% human serum. 50 inhibits the activation of C4.

[0019]

[0023] In some embodiments, the antibodies or antigen-binding fragments provided herein have a K in the range of 0.01 nM to 1 nM as measured by SPR. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to human MASP-2 with a K in the range of 0.05 nM to 0.5 nM as measured by SPR. D It binds to human MASP-2.

[0020]

[0024] Also provided herein are polynucleotides encoding the antibodies or antigen-binding fragments described herein. Also provided herein are vectors comprising the polynucleotides described herein. Also provided herein are host cells comprising the polynucleotides described herein or the vectors described herein.

[0021]

[0025] Also provided herein is a pharmaceutical composition comprising a therapeutically effective amount of an antibody or antigen-binding fragment described herein and a pharmaceutically acceptable carrier.

[0026] Also provided herein is a method for inhibiting MASP-2-dependent complement activation in a subject in need thereof, the method comprising administering to the subject an effective amount of an antibody or antigen-binding fragment described herein. In some embodiments, the subject has a disease or disorder associated with MASP-2-dependent complement activation.

[0022]

[0027] In some embodiments, provided herein are methods for treating a disease or disorder associated with MASP-2-dependent complement activation in a subject in need of such treatment, the method comprising administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment described herein. In some embodiments, the methods provided herein further comprise administering an additional therapy to the subject.

[0023]

[0028] In some embodiments of the methods provided herein, the subject is a human.

[0029] Also provided herein is the use of an antibody or antigen-binding fragment described herein in the treatment of a disease or disorder associated with MASP-2-dependent complement activation.

[0024]

[0030] Also provided herein is the use of an antibody or antigen-binding fragment described herein for the preparation of a medicament for the treatment of a disease or disorder associated with MASP-2-dependent complement activation.

[0025]

[0031] In some embodiments of the methods or uses described herein, the disease or disorder is a kidney disease or disorder, a vascular disease or disorder, a skin disease or disorder, an ophthalmologic disease or disorder, a nervous system disease or disorder, a blood disease or disorder, a musculoskeletal disease or disorder, a genitourinary disease or disorder, a metabolic disease or disorder, an endocrine disease or disorder, a gastrointestinal disease or disorder, or a pulmonary disease or disorder.

[0026]

[0032] In some embodiments, the disease or disorder is IgA nephropathy (IgAN), thrombotic microangiopathy (TMA), lupus nephritis (LN), membranous nephropathy (MN), or C3 glomerulopathy (C3G). In some embodiments, the disease or disorder is IgAN. In some embodiments, the disease or disorder is TMA.

[0027]

[0033] In some embodiments, the disease or disorder is atypical hemolytic uremic syndrome (aHUS), hematopoietic stem cell transplant-associated TMA (HSCT-TMA), thrombotic thrombocytopenic purpura (TTP), TMA secondary to cancer, TMA secondary to chemotherapy, or TMA secondary to transplant. In some embodiments, the disease or disorder is HSCT-TMA. In some embodiments, the disease or disorder is TTP. [Brief explanation of the drawings]

[0028]

[0034] [Figure 1] Figure 1 shows representative results of an in vitro C4 activation assay demonstrating the C4 blocking activity of various MASP-2 antibody clones. Clone 3E10 showed the strongest activity.

[0029]

[0035] [Figure 2] 1 shows representative results of an in vitro C4 activation assay comparing the C4 blocking activity of 3E10 with that of the benchmark antibody, narsoplimab ("MASP-2-BM").

[0030]

[0036] [Figure 3] shows the results of biolayer interferometry (BLI) measuring the binding affinity of 3E10 to human MASP-2.

[0031]

[0037] [Figure 4] Figure 1 shows representative results of an in vitro C3 activation assay comparing the C3 inhibitory activity of 3E10 with that of MASP-2-BM.

[0032]

[0038] [Figure 5] shows representative results of an in vitro MAC activation assay comparing the MAC inhibitory activity of 3E10 with that of MASP-2-BM.

[0033]

[0039] [Figure 6] 1 shows representative results of an in vitro C4 activation assay showing that 3E10 inhibited the activation of C4 in human serum at different concentrations (5%, 25%, and 50%).

[0034]

[0040] [Figure 7] 1 shows representative results of an in vitro C4 activation assay demonstrating the C4 blocking activity of various humanized 3E10s.

[0035]

[0041] [Figure 8] shows representative results of surface plasmon resonance (SPR) measuring the binding affinity of humanized 3E10 for human MASP-2.

[0036]

[0042] [Figure 9] 1 shows representative results of in vitro C4, C3 and MAC activation assays showing that humanized 3E10 inhibited C4, C3 and MAC activation in serum from healthy donors.

[0037]

[0043] [Figure 10] 1 shows representative results of an in vitro MAC activation assay, demonstrating that humanized 3E10 had no effect on either classical or alternative pathway-dependent activation.

[0038]

[0044] [Figure 11]1 shows representative results of a binding competition assay demonstrating that 3E10 and MASP-2-BM did not compete for binding to human MASP-2.

[0039]

[0045] [Figure 12] Figure 1 shows a diagram of the truncated human MASP-2 protein used for epitope mapping.

[0040]

[0046] [Figure 13] Figure 5 shows representative results of binding assays using various truncated human MASP-2 proteins for epitope mapping of humanized 3E10.

[0047] The present disclosure provides novel antibodies, including antigen-binding fragments, that specifically bind to MASP-2 (e.g., human MASP-2). Also disclosed herein are pharmaceutical compositions comprising therapeutically effective amounts of such antibodies or antigen-binding fragments. Further disclosed herein are uses of such pharmaceutical compositions for treating various disorders associated with complement activation (e.g., IgA nephropathy).

[0041]

[0048] Before the present disclosure is further described, it is to be understood that the present disclosure is not limited to particular embodiments described herein, and that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. 5.1 Definition

[0049] Unless otherwise defined herein, scientific and technical terms used in this disclosure shall have the meanings commonly understood by those of ordinary skill in the art. Furthermore, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Overall, the nomenclature used in connection with, and the techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry, and hybridization described herein are those well known and commonly used in the art.

[0042]

[0050] The terms "a" or "an" entity refer to one or more of that entity, for example, "an antibody" is understood to refer to one or more antibodies.

[0043]

[0051] The term "and / or" as used herein should be interpreted as a specific disclosure of each of the two specified features or components, regardless of the presence or absence of the other. Thus, the term "and / or" used herein in phrases such as "A and / or B" is intended to include "A and B," "A or B," "A" (only), and "B" (only). Similarly, the term "and / or" used in phrases such as "A, B, and / or C" is intended to encompass each of the following aspects: A, B, and C; A, B, or C; A or C; A or B; B, or C; A and C; A and B; B and C; A (only); B (only); and C (only).

[0044]

[0052] As used herein, the term "antibody" and its grammatical equivalents refer to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or any combination of the foregoing, through at least one antigen-binding site (wherein the antigen-binding site is located within the variable region of the immunoglobulin molecule). As used herein, the term encompasses intact polyclonal antibodies, intact monoclonal antibodies, single domain antibodies (sdAbs; camelid antibodies, alpaca antibodies), single-chain Fv (scFv) antibodies, heavy chain antibodies (HCAbs), light chain antibodies (LCAbs), multispecific antibodies, bispecific antibodies, monospecific antibodies, monovalent antibodies, and any other modified immunoglobulin molecule containing an antigen-binding site (e.g., dual variable domain immunoglobulin molecules), so long as the antibody exhibits the desired biological activity. Antibodies also include, but are not limited to, murine antibodies, camelid antibodies, chimeric antibodies, humanized antibodies, and human antibodies. Antibodies may be any of the five major classes of immunoglobulins, namely IgA, IgD, IgE, IgG, and IgM, based on the identity of their heavy chain constant domains, designated alpha, delta, epsilon, gamma, and mu, respectively, or their subclasses (isotypes) (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). Unless expressly indicated otherwise, the term "antibody" as used herein includes "antigen-binding fragments" of intact antibodies. As used herein, the term "antigen-binding fragment" refers to portions or fragments of intact antibodies that are the antigen-determining variable regions of the intact antibody. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab'), Fv, linear antibodies, single-chain antibody molecules (e.g., scFv), heavy-chain antibodies (HCAbs), light-chain antibodies (LCAbs), disulfide-linked scFvs (dsscFvs), diabodies, triabodies, tetrabodies, minibodies, dual variable domain antibodies (DVDs), single variable domain antibodies (sdAbs; e.g., camelid antibodies, alpaca antibodies, etc.), and single variable domains of heavy-chain antibodies (VHHs), as well as bispecific or multispecific antibodies formed from antibody fragments.A "bispecific" antibody is an artificial hybrid antibody having two different antigen-binding sites that recognize and specifically bind to two different targets. Bispecific antibodies can be produced by a variety of methods, including fusion of hybridomas or linking of Fab' fragments. See, for example, Songsivilai and Lachmann, Clin. Exp. Immunol. 79:315-321 (1990); Kostelny et al., J. Immunol. 148, 1547-1553 (1992).

[0045]

[0053] The term "humanized antibody," as used herein, refers to forms of non-human (e.g., murine) antibodies that are specific immunoglobulin chains, chimeric immunoglobulins, or fragments thereof that contain minimal non-human sequences. Typically, humanized antibodies are human immunoglobulins. In some cases, residues from the Fv framework regions of a human immunoglobulin are replaced with corresponding residues in an antibody from a non-human species. In some cases, residues from the CDRs are replaced with residues from a CDR of a non-human species (e.g., mouse, rat, hamster, camel) that has the desired specificity, affinity, and / or binding capacity. Humanized antibodies can be further modified by substitution of additional residues either in the Fv framework regions and / or within the replaced non-human residues to improve and optimize the antibody's specificity, affinity, and / or binding capacity. As used herein, the term "human antibody" refers to an antibody having an amino acid sequence corresponding to an antibody produced by a human or prepared using any technique known in the art.

[0046]

[0054] The term "heavy chain," when used in reference to antibodies, refers to a polypeptide chain of approximately 50 to 70 kDa, the amino-terminal portion of which contains a variable region of approximately 120 to 130 or more amino acids, and the carboxy-terminal portion of which contains a constant region. The constant region can be one of five different types (e.g., isotypes), called alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the heavy chain constant region. Different heavy chains vary in size, with α, δ, and γ containing approximately 450 amino acids, while μ and ε contain approximately 550 amino acids. When combined with light chains, these different types of heavy chains give rise to the five well-known classes of antibodies: IgA, IgD, IgE, IgG, and IgM, including the four subclasses of IgG, namely, IgG1, IgG2, IgG3, and IgG4, respectively. The heavy chain can be a human heavy chain.

[0047]

[0055] When used in reference to antibodies, the term "light chain" refers to a polypeptide chain of approximately 25 kDa, the amino-terminal portion of which contains a variable region of about 100 to about 110 or more amino acids, and the carboxy-terminal portion of which contains a constant region. The approximate length of a light chain is 211 to 217 amino acids. Based on the amino acid sequence of the constant domain, there are two different types, called lambda (λ) and kappa (κ). The amino acid sequences of light chains are well known in the art. The light chain may be a human light chain.

[0048]

[0056] The term "variable domain" or "variable region" refers to the portion of an antibody light or heavy chain, usually located at the amino terminus of the light or heavy chain, approximately 120-130 amino acids in length for heavy chains and approximately 100-110 amino acids in length for light chains, and is used in relation to the binding and specificity of each particular antibody to its particular antigen. Variable domains vary significantly in sequence among different antibodies. Sequence variability is concentrated in the CDRs, while the less variable portions of variable domains are called framework regions (FRs). The CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with the antigen. The numbering of amino acid positions used herein is according to the EU index, as found in Kabat et al. (1991), Sequences of Proteins of Immunological Interest (US Department of Health and Human Services, Washington, DC), 5th Edition. The variable region may be a human variable region.

[0049]

[0057] CDR refers to one of the three hypervariable regions (H1, H2, or H3) within the non-framework region of the immunoglobulin (Ig or antibody) VH β-sheet framework or one of the three hypervariable regions (L1, L2, or L3) within the non-framework region of the antibody VL β-sheet framework. Thus, CDRs are variable region sequences interspersed within framework region sequences. CDR regions are well known to those skilled in the art and have been defined by various methods / systems. These systems and / or definitions have been developed and refined over the years and include Kabat, Chothia, IMGT, AbM, and Contact. For example, Kabat defined the most hypervariable regions within antibody variable (V) domains (Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat, Adv. Prot. Chem. 32: 1-75 (1978)). The Chothia definition is based on the location of structural loop regions, which are not part of the conserved β-sheet framework and therefore define CDR region sequences as residues that can adopt various conformations (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987)). Both terminologies are well recognized in the art. Furthermore, the IMGT system is based on sequence variability and location within the structure of the variable region. The AbM definition is a compromise between Kabat and Chothia. The Contact definition is based on analysis of available antibody crystal structures. Software programs (e.g., abYsis) for analyzing antibody sequences and determining CDRs are available and known to those of skill in the art. The locations of CDRs within standard antibody variable domains have been determined by numerous structural comparisons (Al-Lazikani et al., J. Mol. Biol. 273:927-948(1997); Morea et al., Methods 20:267-279(2000)).Because the number of residues within a hypervariable region varies among different antibodies, in standard variable domain numbering schemes, additional residues relative to the standard positions are conventionally numbered a, b, c, etc. next to the residue number (Al-Lazikani et al., supra (1997)). Such nomenclature is also well known to those skilled in the art.

[0050]

[0058] For example, CDRs defined according to either the Kabat (hypervariable) notation or the Chothia (structural) notation are set forth in the table below.

[0051] [Table 1]

[0052]

[0059] One or more CDRs can also be incorporated into a molecule, either covalently or noncovalently, to form an immunoadhesin. Immunoadhesins can incorporate the CDR(s) as part of a larger polypeptide chain, can be covalently linked to another polypeptide chain, or can incorporate the CDR(s) noncovalently. The CDR(s) enable the immunoadhesin to bind to a specific antigen of interest. CDR regions can be analyzed, for example, at the abysis website (http: / / abysis.org / ).

[0053]

[0060] The terms "epitope" and "antigenic determinant" are used interchangeably herein and refer to a site on the surface of a target molecule to which an antibody or antigenic fragment binds, e.g., a localized region on the surface of an antigen. Target molecules can include proteins, peptides, nucleic acids, carbohydrates, or lipids. An epitope with immunogenic activity is a portion of a target molecule that elicits an immune response in an animal. An epitope of a target molecule with antigenic activity is a portion of a target molecule to which an antibody binds, as determined by any method known in the art, including, for example, immunoassays. An antigenic epitope is not necessarily immunogenic. Epitopes are composed of chemically active surface groupings of molecules, such as amino acids or sugar side chains, and often have specific three-dimensional structural characteristics and specific charge characteristics. The term "epitope" includes linear epitopes and conformational epitopes. The region of a target molecule (e.g., a polypeptide) that contributes to an epitope may be contiguous amino acids of the polypeptide, or the epitope may be formed from two or more non-contiguous regions of the target molecule. An epitope may or may not be a three-dimensional surface feature of the target molecule. Epitopes formed from contiguous amino acids (also called linear epitopes) are typically retained upon protein denaturation, whereas epitopes formed by tertiary folding (also called conformational epitopes) are typically lost upon protein denaturation. An epitope typically contains at least three, and more commonly five, six, seven, or eight to ten, amino acids in a unique spatial arrangement.

[0054]

[0061] As used herein, the term "specifically binds" means that a polypeptide or molecule interacts with an epitope, protein, or target molecule more frequently, rapidly, for a longer duration, with greater affinity, or with some combination of the above, than alternative substances, including related and unrelated proteins. Binding moieties (e.g., antibodies) that specifically bind to a target molecule (e.g., an antigen) can be identified, for example, by immunoassay, ELISA, biolayer interferometry ("BLI"), SPR (e.g., Biacore), or other techniques known to those skilled in the art. Typically, a specific response is at least two times the background signal or noise, and may be more than 10 times the background. For a discussion of antibody specificity, see, e.g., Paul (ed.), 1989, Fundamental Immunology, 2nd ed., Raven Press, New York, pp. 332-336. A binding moiety that specifically binds to a target molecule can bind to the target molecule with an affinity that is higher than its affinity for a different molecule. In some embodiments, a binding moiety that specifically binds to a target molecule can bind to the target molecule with an affinity that is at least 20 times greater, at least 30 times greater, at least 40 times greater, at least 50 times greater, at least 60 times greater, at least 70 times greater, at least 80 times greater, at least 90 times greater, or at least 100 times greater than its affinity for a different molecule. In some embodiments, a binding moiety that specifically binds to a particular target molecule binds to a different molecule with such low affinity that binding cannot be detected using assays described herein or otherwise known in the art. In some embodiments, "specifically binds" means, for example, that the binding moiety binds to a different molecule with a K D In some embodiments, "specifically binds" means that the polypeptide or molecule binds to a target molecule with a K of about 10 μM or less, or about 1 μM or less. D In some embodiments, "specifically binds" means that the polypeptide or molecule binds to a target with a K of about 0.1 μM or less, about 0.01 μM or less, or about 1 nM or less.D Specific binding means binding to a target at a specific site. Due to sequence identity between homologous proteins in different species, specific binding may include polypeptides or molecules that recognize proteins or targets in multiple species. Similarly, due to homology within certain regions of the polypeptide sequences of different proteins, specific binding may include polypeptides or molecules that recognize multiple proteins or targets. It is understood that in some embodiments, a binding moiety (e.g., an antibody) that specifically binds to a first target may or may not specifically bind to a second target. Thus, "specific binding" does not necessarily require (although can include) exclusive binding, i.e., binding to a single target. Thus, a binding moiety (e.g., an antibody) can, in some embodiments, specifically bind to multiple targets. For example, in some cases, an antibody contains two identical antigen-binding sites, each capable of specifically binding to the same epitope on two or more proteins. In certain alternative embodiments, an antibody may be bispecific and contain at least two antigen-binding sites with different specificities.

[0055]

[0062] As used herein, the term "binding affinity" typically refers to the strength of the sum of non-covalent interactions between a binding moiety and a target molecule (e.g., an antigen). Binding between a binding moiety and a target molecule is a reversible process, and the affinity of binding is typically measured by the equilibrium dissociation constant (K D ) is reported as K D is the dissociation rate (k off or k d ) and association rate (k on or k a ) is the ratio of the K D The lower the K, the higher the affinity. Various methods for measuring binding affinity are known in the art, any of which may be used for purposes of the present disclosure. Specific exemplary embodiments include the following: In some embodiments, "K" is used to measure binding affinity. D " or "K D The "K value" can be measured by assays known in the art, for example, by binding assays. Dcan be measured by radiolabeled antigen binding assay (RIA) (Chen et al., (1999) J. Mol Biol 293:865-881). D or K D K values can also be measured by using biolayer interferometry (BLI), for example, using the Gator system (Probe Life) or the Octet-96 system (Sartorius AG). D or K D Values can also be measured by using a surface plasmon resonance assay by Biacore, for example, using a BIAcore™-2000 or BIAcore™-3000 (BIAcore, Inc., Piscataway, NJ).

[0056]

[0063] The term "variant," as used herein in reference to a protein or polypeptide having particular sequence characteristics ("reference polypeptide" or "reference polypeptide"), refers to a different protein or polypeptide that has one or more (e.g., about 1 to about 25, about 1 to about 20, about 1 to about 15, about 1 to about 10, or about 1 to about 5) amino acid substitutions, deletions, and / or additions when compared to the reference protein or polypeptide. The changes to the amino acid sequence can be amino acid substitutions. The changes to the amino acid sequence can be conservative amino acid substitutions. A functional fragment or functional variant of a protein or polypeptide maintains essential structural and functional properties of the reference protein or polypeptide.

[0057]

[0064] The terms "polypeptide," "peptide," and "protein," and their grammatical equivalents, used interchangeably herein, refer to polymers of amino acids of any length, which may be linear or branched, which may include non-naturally occurring amino acids, modified amino acids, or may be interrupted by non-amino acids. Polypeptides, peptides, or proteins may also be modified, for example, by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification.

[0058]

[0065] The terms "polynucleotide," "nucleic acid," and grammatical equivalents, used interchangeably herein, refer to a polymer of nucleotides of any length, including DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase.

[0059]

[0066] The terms "identical," percent "identity," and their grammatical equivalents, when used in the context of two or more polynucleotides or polypeptides, refer to two or more sequences or subsequences that are the same or have a specified percentage of the same nucleotide or amino acid residues when compared and aligned for maximum correspondence (introducing gaps, if necessary), not considering conservative amino acid substitutions as part of the sequence identity. Percent identity can be measured using sequence comparison software or algorithms or by visual inspection. Various algorithms and software that can be used to align amino acid or nucleotide sequences are well known in the art. These include, but are not limited to, BLAST, ALIGN, Megalign, BestFit, GCG Wisconsin Package, and variants thereof. In some embodiments, two polynucleotides or polypeptides provided herein are substantially identical, meaning that they share 70%, at least 75%, at least 80%, at least 85%, at least 90%, and in some embodiments, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% nucleotide or amino acid residue identity when compared and aligned for maximum correspondence, as determined using a sequence comparison algorithm or by visual inspection. In some embodiments, identity exists over a region of the amino acid sequence that is at least about 10 residues, at least about 20 residues, at least about 40-60 residues, at least about 60-80 residues, or any integer value therebetween. In some embodiments, identity exists over a region longer than 60-80 residues, e.g., about 80-100 residues, and in some embodiments, the sequences are substantially identical over the entire length of the sequences being compared, e.g., the coding regions of the target proteins or antibodies. In some embodiments, the identity exists over a region of the nucleotide sequence that is at least about 10 bases, at least about 20 bases, at least about 40-60 bases, at least about 60-80 bases in length, or any integer value therebetween.In some embodiments, the identity exists over a region longer than 60-80 bases, e.g., over at least about 80-1000 bases or more, and in some embodiments, the sequences are substantially identical over the entire length of the sequence being compared, e.g., the nucleotide sequence encoding a protein of interest.

[0060]

[0067] As used herein, the term "vector" and its grammatical equivalents refer to a vehicle used to transport genetic material (e.g., a polynucleotide sequence) that can be introduced into a host cell and replicated and / or expressed therein. Vectors applicable for use include, for example, expression vectors, plasmids, phage vectors, viral vectors, episomes, artificial chromosomes, and the like, which may contain selectable sequences or markers operable for stable integration into a host cell chromosome. Furthermore, a vector may contain one or more selectable marker genes and appropriate expression control sequences. Selectable marker genes that may be included provide, for example, resistance to antibiotics or toxins, complement auxotrophic deficiencies, or supply critical nutrients not present in the culture medium. Expression control sequences may include constitutive and inducible promoters, transcription enhancers, transcription terminators, and the like, all of which are well known in the art. When two or more polynucleotides are coexpressed, both polynucleotides may be inserted, for example, into a single expression vector or into separate expression vectors. In the case of single vector expression, the encoding polynucleotides may be operably linked to one common expression control sequence, or may be linked to different expression control sequences, such as one inducible promoter and one constitutive promoter. Introduction of the polynucleotide into the host cell can be confirmed using methods well known in the art. It will be understood by those skilled in the art that the polynucleotide will be expressed in an amount sufficient to produce the desired product (e.g., an anti-MASP-2 antibody or antigen-binding fragment described herein), and further that the expression level can be optimized to obtain sufficient expression using methods well known in the art.

[0061]

[0068] As used herein, the term "encode" and its grammatical equivalents refer to the inherent property of a particular sequence of nucleotides in a polynucleotide or nucleic acid, such as a gene, cDNA, or mRNA, to serve as a template for the synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA, and mRNA) or a defined sequence of amino acids, and the biological property resulting therefrom. Thus, a gene encodes a protein when transcription and translation of the mRNA corresponding to that gene produces the protein. Unless otherwise specified, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate versions of each other and encode the same amino acid sequence. Nucleotide sequences that encode proteins and RNAs may contain introns.

[0062]

[0069] An "isolated" polypeptide, peptide, protein, antibody, polynucleotide, vector, cell, or composition is a polypeptide, peptide, protein, antibody, polynucleotide, vector, cell, or composition in a form not found in nature. Isolated polypeptides, peptides, proteins, antibodies, polynucleotides, vectors, cells, or compositions include those that have been purified to the extent that they are no longer in a form found in nature. In some embodiments, an isolated polypeptide, peptide, protein, antibody, polynucleotide, vector, cell, or composition is substantially pure.

[0063]

[0070] The term "treating" and its grammatical equivalents as used herein in reference to a disease or condition, or a subject having a disease or condition, refers to the act of suppressing, eliminating, reducing, and / or ameliorating the symptoms, the severity of the symptoms, and / or the frequency of the symptoms associated with the disease or disorder being treated.

[0064]

[0071] As used herein, the term "administering" and its grammatical equivalents refer to the act of delivering or causing to be delivered a therapeutic agent or pharmaceutical composition to the body of a subject, by methods described herein or otherwise known in the art. A therapeutic agent can be a compound, a polypeptide, an antibody, a cell, or a population of cells. Administering a therapeutic agent or pharmaceutical composition includes formulating the therapeutic agent or pharmaceutical composition to be delivered to the body of a subject. Exemplary dosage forms include oral dosage forms such as tablets, capsules, syrups, suspensions, etc.; injectable dosage forms such as intravenous (IV), intramuscular (IM), or intraperitoneal (IP); transdermal dosage forms such as creams, jellies, powders, patches, etc.; buccal dosage forms; inhalation powders, sprays, suspensions, and rectal suppositories.

[0065]

[0072] As used herein, the terms "effective amount," "therapeutically effective amount," and their grammatical equivalents refer to the administration of an agent to a subject, either alone or as part of a pharmaceutical composition, in an amount that, when administered to a subject, is capable of having any detectable positive effect on any symptom, aspect, or characteristic of a disease, disorder, or condition, either in a single dose or as part of a series of doses. A therapeutically effective amount can be ascertained by measuring the relevant physiological effect. The precise amount needed will vary from subject to subject, depending on the subject's age, weight, and general condition, the severity of the condition being treated, the judgment of the clinician, and the like. An appropriate "effective amount" in any individual case can be determined by one of ordinary skill in the art using routine experimentation.

[0066]

[0073] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refers to a material that is suitable for administering a drug to an individual together with an active agent without causing any undesired biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition.

[0067]

[0074] As used herein, the term "subject" refers to any animal (e.g., mammal) that will be the recipient of a particular treatment, including, but not limited to, humans, non-human primates, dogs, cats, rodents, etc. The subject may be a human. The subject may have a particular disease or condition.

[0068]

[0075] Ranges: Throughout this disclosure, various aspects of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the broadness of the range.

[0069]

[0076] Exemplary genes and polypeptides are described herein by reference to GenBank numbers, GI numbers, and / or SEQ ID NOs. It is understood that one of skill in the art can readily identify homologous sequences by reference to sequence sources, including, but not limited to, GenBank (ncbi.nlm.nih.gov / genbank / ) and EMBL (embl.org / ). 5.2 Lectin pathway and MASP-2

[0077] The complement system (CS) is an essential part of innate immunity. It is a network of over 30 plasma and cell surface proteins that recognize, mark, and remove microbial pathogens and dangerously altered (e.g., apoptotic) self-cells, inducing inflammation, and recruiting immune cells. The CS can be activated by three pathways. The classical pathway is primarily activated by immune complexes but can also recognize microbial surfaces, apoptotic, and necrotic cells. It contributes to the removal of unnecessary synapses during ontogeny and is important for the clearance of immune complexes and cellular debris. The lectin pathway recognizes ancient, surface-exposed molecular determinants on microorganisms via a variety of pattern recognition molecules (PRMs) and provides instantaneous defense against microbial pathogens without relying on specific antibodies. The alternative pathway (AP) continuously attacks all surfaces through low-level spontaneous activation but is productively activated only on surfaces lacking protective complement regulatory molecules. Furthermore, the AP provides an important amplification loop for complement activation.

[0070]

[0078] Danger signal recognition triggers the activation of pathway-specific serine proteinase zymogens. The activated proteinases cleave downstream complement components to form surface-bound C3 convertases: C4b2a for the classical and lectin pathways, and C3bBb for the alternative pathway. At this point, the three activation pathways converge onto a common effector pathway, resulting in the targeting and lysis of pathogens, the recruitment of immune cells, and the induction of inflammation.

[0071]

[0079] Normally, complement activation is tightly regulated. Lack of complement inhibition is the underlying pathological mechanism for many clinical conditions. In most complement-related diseases, the contribution of one of three pathways is dominant. Pathway-specific inhibitors, such as inhibitors of pathway-specific proteinases, could be ideal therapeutic agents that selectively block the disrupted pathological pathway while sparing the protective functions of the other two pathways.

[0072]

[0080] Currently, three mannan-binding lectin-associated serine proteinases (MASP-1, MASP-2, and MASP-3) are known to associate with mannan-binding lectin (MBL) (also known as "mannose-binding protein") in human serum. The MBL-MASP-2 complex plays an important role in innate immunity through MBL binding to carbohydrate structures present on diverse microorganisms. MASP-2 is activated when the recognition components bind to their respective patterns and can also be activated by MASP-1, subsequently cleaving complement component C4 into C4a and C4b. After the cleavage product C4b binds to plasma C2, C4b-bound C2 becomes a substrate for a second MASP-2-mediated cleavage step that converts C4b-bound C2 into the enzymatically active complex C4bC2a and small C2b cleavage fragments. C4b2a is the C3 convertase of the lectin pathway, converting the abundant plasma component C3 to C3a and C3b. C3b binds to any nearby surface via a thioester bond. When several C3b fragments bind in close proximity to the C3 convertase complex C4b2a, this convertase changes its specificity to convert C5 to C5b and C5a, forming the C5 convertase complex C4b2a(C3b)n. This C5 convertase can initiate the formation of the membrane attack complex (MAC, a complex of five terminal complement components (C5b combined with C6, C7, C8, and C9) (also called C5b-9)) that inserts into and disrupts membranes, but this process alone is thought to be insufficient to promote lysis. Rather, the initial C3b opsonin produced by the lectin pathway nucleates the formation of new alternative pathway C3 convertase and C5 convertase sites, which ultimately result in abundant MAC formation and lysis. There is also a MASP-2-dependent C4 bypass activation pathway to activate C3 in the absence of C4, which plays an important role in the pathophysiology of ischemia-reperfusion injury.

[0073]

[0081] Thus, as used herein and as understood in the art, "MASP-2-dependent complement activation" refers to the lectin pathway of complement activation, which requires MASP-2. MASP-2-dependent complement activation occurs in the presence of Ca++, resulting in the formation of the lectin pathway C3 convertase C4b2a, and the accumulation of the C3 cleavage product C3b, which can then generate the C5 convertase C4b2a (C3b)n, which can trigger opsonization and / or lysis.

[0074]

[0082] The human MASP-2 gene is located on chromosome 1p36.3-2 (Stover et al., Cytogenet and Cell Genet 84: 148-149 (1999)) and encompasses 12 exons. Human MASP-2 cDNA is encoded by exons 2, 3, 4, 6, 7, 8, 9, 10, 11, and 12. A 20 kDa protein called MBL-associated protein 19 (also called "MAp19" or "sMAP") arises from exons 2, 3, 4, and 5. MAp19 is a non-enzymatic protein containing the N-terminal CUB1-EGF region of MASP-2 with four additional residues (EQSL) derived from exon 5.

[0075]

[0083] The MASP-2 polypeptide has 686 amino acid residues, including a 15-residue leader peptide that is cleaved after secretion to yield the mature form of human MASP-2 (671 amino acids, SEQ ID NO: 1). The MASP-2 polypeptide exhibits a molecular structure similar to that of MASP-1, MASP-3, and the proteinases C1r and C1s of the C1 complement system. A schematic diagram showing the domain structure of the human MASP-2 polypeptide is shown in Figure 12 (labeled "hMASP2-FL"). As shown in Figure 12, the serine proteinase MASP-2 consists of six distinct domains: (1) an N-terminal C1r / C1 / sea urchin VEGF / bone morphogenetic protein (or CUB1) domain (aa 1-122 of SEQ ID NO: 1), (2) an epidermal growth factor (EGF)-like domain (aa 123-166 of SEQ ID NO: 1), (3) a second CUB domain (CUB2) (aa 167-279 of SEQ ID NO: 1), (4 and 5) two complement control protein (CCP1 and CCP2) domains (CCP1 aa 280-345 and CCP2 aa 346-414 of SEQ ID NO: 1), and (6) a serine proteinase (SP) domain (aa 415-671 of SEQ ID NO: 1). (SEQ ID NO: 1)

[0084] The MASP-2 polypeptide has an α chain (heavy chain) containing the CUB1-EGF-CUB2-CCP1-CCP2 domains and a β chain (light chain) containing a serine protease domain. The CUBl, EGF, and CUB2 domains are required for dimerization, and the CUBl, EGF, CUB2, and CCP1 domains contain the binding site for MBP. Each MASP-2 dimer binds to two MBL subunits (Wallis et al., J. Biol Chem. 279:14065-14073 (2004)). 5.3 Anti-MASP-2 antibodies and antigen-binding fragments

[0085] Provided herein are antibodies or antigen-binding fragments thereof that specifically bind to MASP-2 (e.g., human MASP-2). In some embodiments, anti-MASP-2 antibodies are provided. In some embodiments, the antibody is an IgA, IgD, IgE, IgG, or IgM antibody. In some embodiments, the antibody is an IgA antibody. In some embodiments, the antibody is an IgD antibody. In some embodiments, the antibody is an IgE antibody. In some embodiments, the antibody is an IgG antibody. In some embodiments, the antibody is an IgM antibody. In some embodiments, the antibodies provided herein may be an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the antibody is an IgG1 antibody. In some embodiments, the antibody is an IgG2 antibody. In some embodiments, the antibody is an IgG3 antibody. In some embodiments, the antibody is an IgG4 antibody.

[0076]

[0086] In some embodiments, antigen-binding fragments of anti-MASP-2 antibodies are provided. In some embodiments, the antigen-binding fragments provided herein may be single domain antibodies (sdAbs), heavy chain antibodies (HCAbs), Fabs, Fab's, F(ab')2, Fvs, single-chain variable fragments (scFvs), or (scFv)2s. In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are single domain antibodies (sdAbs). In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are heavy chain antibodies (HCAbs). In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are Fabs. In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are Fab's. In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are F(ab')2s. In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are Fvs. In some embodiments, the antigen-binding fragments of anti-MASP-2 antibodies are scFvs. In some embodiments, the antigen-binding fragment of the anti-MASP-2 antibody is a disulfide-linked scFv [(scFv)2]. In some embodiments, the antigen-binding fragment of the anti-MASP-2 antibody is a diabody (dAb).

[0077]

[0087] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise recombinant antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise monoclonal antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise polyclonal antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise Camelidae (e.g., camel, dromedary, and llama) antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise chimeric antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise humanized antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise human antibodies or antigen-binding fragments. In some embodiments, an anti-MASP-2 human scFv is provided.

[0078]

[0088] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein are isolated. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein are substantially pure.

[0079]

[0089] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise multispecific antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise bispecific antibodies or antigen-binding fragments. In some embodiments, the bispecific antibodies or antigen-binding fragments comprise anti-MASP-2 antibodies or antigen-binding fragments provided herein. In some embodiments, the bispecific antibodies or antigen-binding fragments comprise anti-MASP-2 scFvs provided herein.

[0080]

[0090] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise a monovalent antigen-binding site. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments comprise a monospecific binding site. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments comprise a bivalent binding site.

[0081]

[0091] In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment is a monoclonal antibody or antigen-binding fragment. Monoclonal antibodies can be prepared by any method known to those skilled in the art. One exemplary approach is screening a protein expression library, such as a phage or ribosome display library. Phage display is described, for example, in U.S. Patent No. 5,223,409 by Ladner et al.; Smith (1985) Science 228:1315-1317; and WO 92 / 18619. In some embodiments, recombinant monoclonal antibodies are isolated from a phage display library expressing the variable regions or CDRs of the desired species. Screening of phage libraries can be achieved by various techniques known in the art.

[0082]

[0092] In some embodiments, monoclonal antibodies are prepared using hybridoma methods known to those skilled in the art. For example, using the hybridoma method, a mouse, rat, rabbit, hamster, or other suitable host animal is immunized as described above. In some embodiments, lymphocytes are immunized in vitro. In some embodiments, the immunizing antigen is a human protein or fragment thereof. In some embodiments, the immunizing antigen is a human protein or fragment thereof.

[0083]

[0093] After immunization, lymphocytes are isolated and fused with a suitable myeloma cell line, e.g., using polyethylene glycol. Hybridoma cells are selected using specialized media known in the art; unfused lymphocytes and myeloma cells do not survive the selection process. Hybridomas producing monoclonal antibodies against the selected antigen can be identified by various methods, including, but not limited to, immunoprecipitation, immunoblotting, and in vitro binding assays (e.g., flow cytometry, FACS, ELISA, BLI, SPR (e.g., Biacore), and radioimmunoassay). Once hybridoma cells producing antibodies of the desired specificity, affinity, and / or activity are identified, clones can be subcloned by limiting dilution or other techniques. Hybridomas can be grown in vitro in culture using standard methods or in vivo as ascites tumors in animals. Monoclonal antibodies can be purified from the culture medium or ascites fluid according to standard methods in the art, including, but not limited to, affinity chromatography, ion exchange chromatography, gel electrophoresis, and dialysis.

[0084]

[0094] In some embodiments, monoclonal antibodies are produced using recombinant DNA techniques known to those skilled in the art. For example, polynucleotides encoding the antibodies are isolated from mature B cells or hybridoma cells, e.g., by RT-PCR using oligonucleotide primers that specifically amplify genes encoding the antibody heavy and light chains, and their sequences are determined by standard techniques. The isolated polynucleotides encoding the heavy and light chains are then cloned into suitable expression vectors that produce the monoclonal antibodies when transfected into host cells that do not otherwise produce immunoglobulin proteins, such as E. coli, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells.

[0085]

[0095] In some embodiments, monoclonal antibodies are modified using recombinant DNA technology to generate surrogate antibodies. In some embodiments, the light and heavy chain constant domains of a murine monoclonal antibody are replaced with constant regions from a human antibody to generate chimeric antibodies. In some embodiments, the constant regions are truncated or removed to generate desired antibody fragments of the monoclonal antibody. In some embodiments, site-directed or high-density mutagenesis of the variable regions is used to optimize the specificity and / or affinity of the monoclonal antibody.

[0086]

[0096] In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment is a humanized antibody or antigen-binding fragment. Various methods for producing humanized antibodies are known in the art. Methods for achieving high-affinity binding with humanized antibodies are known in the art. A non-limiting example of such a method is hypermutation of the variable region and selection of cells expressing such high-affinity antibodies (affinity maturation). In addition to using a display library, a specific antigen (e.g., recombinant MASP-2 or an epitope thereof) can be used to immunize a non-human animal, e.g., a rodent. In certain embodiments, rodent antigen-binding fragments (e.g., murine antigen-binding fragments) can be produced and isolated using methods known in the art and / or disclosed herein. In some embodiments, a mouse can be immunized with an antigen (e.g., recombinant MASP-2 or an epitope thereof).

[0087]

[0097] In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment is a human antibody or antigen-binding fragment. Human antibodies can be prepared using various techniques known in the art. In some embodiments, human antibodies are produced from immortalized human B lymphocytes immunized in vitro. In some embodiments, human antibodies are produced from lymphocytes isolated from immunized individuals. In either case, cells producing antibodies against the target antigen can be generated and isolated. In some embodiments, human antibodies are selected from phage libraries, where the phage library expresses human antibodies. Alternatively, phage display technology can be used to generate human antibodies and antibody fragments in vitro from immunoglobulin variable region gene repertoires derived from unimmunized donors. Techniques for generating and using antibody phage libraries are well known in the art. Once an antibody is identified, affinity maturation strategies known in the art, including, but not limited to, chain shuffling and site-directed mutagenesis, can be used to generate higher affinity human antibodies. In some embodiments, human antibodies are produced in transgenic mice containing human immunoglobulin loci. Upon immunization, these mice are capable of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production.

[0088]

[0098] In some embodiments, anti-MASP-2 antibody clone 3E10 is provided. Its sequence characteristics are described below. The specific CDR sequences defined herein are generally based on either the Kabat or IMGT definitions. However, it is understood that reference to the heavy chain CDR(s) and / or light chain CDR(s) of a specific antibody encompasses all CDR definitions known to those skilled in the art.

[0089] [Table 2]

[0090] [Table 3]

[0091]

[0099] CDR grafting was performed according to the Kabat numbering to generate humanized 3E10. Thus, the Kabat CDRs remained unchanged after humanization, but some IMGT CDRs showed minor changes (1-3 amino acid substitutions).

[0092]

[0100] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise one, two, three, four, five, and / or six CDRs of any one of the antibodies described herein. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise a light chain variable region (VL) comprising one, two, and / or three light chain CDRs (VL CDRs) in Table 1. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise a heavy chain variable region (VH) comprising one, two, and / or three heavy chain CDRs (VH CDRs) in Table 2. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein comprise one, two, and / or three VL CDRs in Table 1 and one, two, and / or three VH CDRs in Table 2.

[0093]

[0101] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 comprises a VL comprising (1) a VL CDR1 having the amino acid sequence of SEQ ID NO: 7 or 8, (2) a VL CDR2 having the amino acid sequence of SEQ ID NO: 9 or 10, or (3) a VL CDR3 having the amino acid sequence of SEQ ID NO: 11, or a variant thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VL CDR; and / or (1) a VH CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 12 to 15, (2) a VH CDR2 having the amino acid sequence of SEQ ID NO: 16 or 17, or (3) a VH CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 18 to 20, or the VH Antibodies or antigen-binding fragments thereof comprising a VH are provided, including these variants having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the CDRs.

[0094]

[0102] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL comprising (1) a VL CDR1 having the amino acid sequence of SEQ ID NO: 7 or 8, (2) a VL CDR2 having the amino acid sequence of SEQ ID NO: 9 or 10, or (3) a VL CDR3 having the amino acid sequence of SEQ ID NO: 11, or a variant thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VL CDR. In some embodiments, the variant has about 5 amino acid substitutions, additions, and / or deletions in the VL CDR.

[0095]

[0103] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL comprising (1) a VL CDR1 having the amino acid sequence of SEQ ID NO: 7 or 8, (2) a VL CDR2 having the amino acid sequence of SEQ ID NO: 9 or 10, and (3) a VL CDR3 having the amino acid sequence of SEQ ID NO: 11, or a variant thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VL CDR. In some embodiments, the variant has up to about 5 amino acid substitutions, additions, and / or deletions in the VL CDR.

[0096]

[0104] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, wherein the VL comprises VL CDR1, CDR2 and CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9 and 11, respectively, as defined by Kabat, or variants thereof having substitutions, additions and / or deletions of up to about 3, about 5, about 8, about 10, about 12 or about 15 amino acids in the VL CDR. In some embodiments, the variants have substitutions, additions and / or deletions of up to about 5 amino acids in the VL CDR. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, which has a VL, wherein the VL comprises VL CDR1, CDR2 and CDR3 having the amino acid sequences of SEQ ID NOs: 8, 10 and 11, respectively, as defined by IMGT, or variants thereof having substitutions, additions and / or deletions of up to about 3, about 5, about 8, about 10, about 12 or about 15 amino acids in the VL CDR. In some embodiments, the variants have substitutions, additions and / or deletions of up to about 5 amino acids in the VL CDR.

[0097]

[0105] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH comprising (1) a VH CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 12 to 15, (2) a VH CDR2 having an amino acid sequence of SEQ ID NO: 16 or 17, or (3) a VH CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 18 to 20, or a variant thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VH CDR. In some embodiments, the variant has up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDR.

[0098]

[0106] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH comprising (1) a VH CDR1 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 12 to 15, (2) a VH CDR2 having an amino acid sequence of SEQ ID NO: 16 or 17, and (3) a VH CDR3 having an amino acid sequence selected from the group consisting of SEQ ID NOs: 18 to 20, or a variant thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VH CDR. In some embodiments, the variant has up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDR.

[0099]

[0107] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, having a VH, wherein the VH comprises VH CDR1, CDR2 and CDR3 having the amino acid sequences of SEQ ID NOs: 12, 16 and 18, respectively, as defined by Kabat, or variants thereof having substitutions, additions and / or deletions of up to about 3, about 5, about 8, about 10, about 12 or about 15 amino acids in the VH CDRs. In some embodiments, the variants have substitutions, additions and / or deletions of up to about 5 amino acids in the VH CDRs.

[0100]

[0108] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, wherein the VH has a VH CDR1, CDR2, and CDR3 having the amino acid sequence of (1) SEQ ID NOs: 13, 17, and 19, respectively, (2) SEQ ID NOs: 14, 17, and 19, respectively, (3) SEQ ID NOs: 15, 17, and 19, respectively, (4) SEQ ID NOs: 13, 17, and 20, respectively, (5) SEQ ID NOs: 14, 17, and 20, respectively, or (6) SEQ ID NOs: 15, 17, and 20, respectively, as defined by IMGT, or a variant thereof having substitutions, additions, and / or deletions of up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acids in the VH CDR. In some embodiments, the variant has substitutions, additions, and / or deletions of up to about 5 amino acids in the VH CDR.

[0101]

[0109] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, which comprises, as defined by Kabat, (a) a VL comprising VL CDR1, VL CDR2 and VL CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9 and 11, respectively, or variants thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VL CDRs, and / or (b) a VH comprising VH CDR1, VH CDR2 and VH CDR3 having the amino acid sequences of SEQ ID NOs: 12, 16 and 18, respectively, or variants thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VH CDRs.

[0102]

[0110] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, which comprises VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2 and VH CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9, 11, 12, 16 and 18, respectively, as defined by Kabat, or variants thereof having substitutions, additions and / or deletions of up to about 3, about 5, about 8, about 10, about 12 or about 15 amino acids in the CDRs.

[0103]

[0111] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, which comprises, as defined by IMGT, (a) a VL comprising VL CDR1, VL CDR2 and VL CDR3 having the amino acid sequences of SEQ ID NOs: 8, 10 and 11, respectively, or variants thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VL CDRs, and / or (b) a VH comprising VH CDR1, VH CDR2 and VH CDR3 having the amino acid sequences of SEQ ID NOs: 13, 17 and 19, respectively, or variants thereof having up to about 3, about 5, about 8, about 10, about 12, or about 15 amino acid substitutions, additions, and / or deletions in the VH CDRs.

[0104]

[0112] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, which, as defined by IMGT, comprises a VL CDR1 having the amino acid sequence of SEQ ID NO: 8, a VL CDR2 having the amino acid sequence of SEQ ID NO: 10, a VL CDR3 having the amino acid sequence of SEQ ID NO: 11, a VH CDR1 having the amino acid sequence of SEQ ID NO: 13, 14 or 15, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 19 or 20.

[0105]

[0113] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL CDR1, a VL CDR2, a VL CDR3, a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 8, 10, 11, 13, 17, and 19, as defined by IMGT. In some embodiments, the VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and VH CDR3 may have SEQ ID NOs: 8, 10, 11, 14, 17, and 19, respectively, as defined by IMGT. In some embodiments, the VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and VH CDR3 may have SEQ ID NOs: 8, 10, 11, 15, 17, and 19, respectively, as defined by IMGT. In some embodiments, the VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and VH CDR3, as defined by IMGT, can have SEQ ID NOs: 8, 10, 11, 13, 17, and 20, respectively. In some embodiments, the VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and VH CDR3, as defined by IMGT, can have SEQ ID NOs: 8, 10, 11, 14, 17, and 20, respectively. In some embodiments, the VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and VH CDR3, as defined by IMGT, can have SEQ ID NOs: 8, 10, 11, 15, 17, and 19, respectively.

[0106] [Table 4-1]

[0107] [Table 4-2]

[0108]

[0114] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL that has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 21. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH that has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 22.

[0109]

[0115] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising: (a) a VL having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 21; and (b) a VH having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 22. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL and a VH, wherein the VL and VH have the amino acid sequences of SEQ ID NOs: 21 and 22, respectively.

[0110]

[0116] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL, wherein the VL has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 21. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VL that has at least 85% sequence identity to SEQ ID NO: 21. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VL that has at least 90% sequence identity to SEQ ID NO: 21. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VL that has at least 95% sequence identity to SEQ ID NO: 21. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VL with at least 98% sequence identity to SEQ ID NO: 21. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL having the amino acid sequence of SEQ ID NO: 21.

[0111]

[0117] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 22. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VH that has at least 85% sequence identity to SEQ ID NO: 22. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VH that has at least 90% sequence identity to SEQ ID NO: 22. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VH that has at least 95% sequence identity to SEQ ID NO: 22. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof has a VH with at least 98% sequence identity to SEQ ID NO: 22. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 22.

[0112]

[0118] In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof comprises a humanized antibody or antigen-binding fragment. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof comprises a VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and / or VH CDR3 derived from an antibody or antigen-binding fragment described herein. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof comprises a variant of the anti-MASP-2 antibody or antigen-binding fragment described herein. A variant of the anti-MASP-2 antibody or antigen-binding fragment may comprise substitutions, additions, and / or deletions of 1 to 30 amino acids in the anti-MASP-2 antibody or antigen-binding fragment. A variant of the anti-MASP-2 antibody or antigen-binding fragment may comprise substitutions, additions, and / or deletions of 1 to 25 amino acids in the anti-MASP-2 antibody or antigen-binding fragment. In some embodiments, the variant of an anti-MASP-2 antibody or antigen-binding fragment comprises 1 to 20 substitutions, additions, and / or deletions in the anti-MASP-2 antibody or antigen-binding fragment. In some embodiments, the variant of an anti-MASP-2 antibody or antigen-binding fragment comprises 1 to 15 substitutions, additions, and / or deletions in the anti-MASP-2 antibody or antigen-binding fragment. In some embodiments, the variant of an anti-MASP-2 antibody or antigen-binding fragment comprises 1 to 10 substitutions, additions, and / or deletions in the anti-MASP-2 antibody or antigen-binding fragment. In some embodiments, the variant of an anti-MASP-2 antibody or antigen-binding fragment comprises 1 to 5 conservative amino acid substitutions, additions, and / or deletions in the anti-MASP-2 antibody or antigen-binding fragment. In some embodiments, the variant of an anti-MASP-2 antibody or antigen-binding fragment comprises 1 to 3 amino acid substitutions, additions, and / or deletions in the anti-MASP-2 antibody or antigen-binding fragment. In some embodiments, the amino acid substitutions, additions, and / or deletions are conservative amino acid substitutions. In some embodiments, the conservative amino acid substitution(s) are present in a CDR of the antibody or antigen-binding fragment. In some embodiments, the conservative amino acid substitution(s) are not present in a CDR of the antibody or antigen-binding fragment.In some embodiments, the conservative amino acid substitution(s) are within framework regions of the antibody or antigen-binding fragment.

[0113]

[0119] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising: (a) a VL having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 23; and (b) a VH having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 24. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL and a VH, wherein the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 24, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 25, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 26, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 27, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 28, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 29, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 30, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 31, respectively. In some embodiments, the VL and VH have the amino acid sequences of SEQ ID NOs: 23 and 32, respectively.

[0114]

[0120] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL, wherein the VL has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 23. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VL that has at least 85% sequence identity to SEQ ID NO: 23. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VL that has at least 90% sequence identity to SEQ ID NO: 23. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VL that has at least 95% sequence identity to SEQ ID NO: 23. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VL having at least 98% sequence identity with SEQ ID NO: 23. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VL having the amino acid sequence of SEQ ID NO: 23.

[0115]

[0121] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 24. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 24. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 24. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 24. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VH having at least 98% sequence identity with SEQ ID NO: 24. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 24.

[0116]

[0122] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 25. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 25. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 25. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 25. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH having at least 98% sequence identity with SEQ ID NO: 25. In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 25.

[0117]

[0123] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 26. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 26. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 26. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 26. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VH having at least 98% sequence identity with SEQ ID NO: 26. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 26.

[0118]

[0124] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 27. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 27. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 27. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 27. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VH with at least 98% sequence identity to SEQ ID NO: 27. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 27.

[0119]

[0125] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 28. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 28. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 28. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 28. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VH having at least 98% sequence identity with SEQ ID NO: 28. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 28.

[0120]

[0126] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 29. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 29. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 29. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 29. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VH having at least 98% sequence identity to SEQ ID NO: 29. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 29.

[0121]

[0127] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 30. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 30. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 30. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 30. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH having at least 98% sequence identity with SEQ ID NO: 30. In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 30.

[0122]

[0128] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 31. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 31. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 31. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 31. The humanized anti-MASP-2 antibody or its antigen-binding fragment may have a VH with at least 98% sequence identity to SEQ ID NO: 31. In some embodiments, a humanized antibody or its antigen-binding fragment that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 31.

[0123]

[0129] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH has at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 32. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 85% sequence identity to SEQ ID NO: 32. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 90% sequence identity to SEQ ID NO: 32. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH that has at least 95% sequence identity to SEQ ID NO: 32. The humanized anti-MASP-2 antibody or antigen-binding fragment thereof may have a VH having at least 98% sequence identity with SEQ ID NO: 32. In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH having the amino acid sequence of SEQ ID NO: 32.

[0124]

[0130] An anti-MASP-2 antibody or antigen-binding fragment thereof may comprise a combination of any VL disclosed herein and any VH disclosed herein. In some embodiments, the VL and VH are linked by a linker. The linker may be a flexible linker or a rigid linker. In some embodiments, the linker has the amino acid sequence (GGGGS)n, n=1, 2, 3, 4, or 5 (SEQ ID NO: 77). In some embodiments, the linker has the amino acid sequence (EAAAK)n, n=1, 2, 3, 4, or 5 (SEQ ID NO: 78). In some embodiments, the linker has the amino acid sequence (PA)nP, n=1, 2, 3, 4, or 5 (SEQ ID NO: 79).

[0125]

[0131] In some embodiments, an antibody or antigen-binding fragment thereof is provided, comprising: (a) a VL comprising VL CDRs 1, 2, and 3 derived from a VL having the amino acid sequence of SEQ ID NO: 21 or 23; and / or (b) a VH comprising VH CDRs 1, 2, and 3 derived from a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 22 and 24 to 32.

[0126]

[0132] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL, wherein the VL comprises VL CDRs 1, 2, and 3 derived from a VL having the amino acid sequence of SEQ ID NO: 21. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL, wherein the VL comprises VL CDRs 1, 2, and 3 derived from a VL having the amino acid sequence of SEQ ID NO: 23.

[0127]

[0133] In some embodiments, a humanized antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 22. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 24. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 25. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 26. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 27. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 28. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 29.In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 30. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 31. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VH, wherein the VH comprises VH CDRs 1, 2, and 3 derived from a VH having the amino acid sequence of SEQ ID NO: 32.

[0128]

[0134] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL and a VH, wherein the VL comprises VL CDR1, CDR2 and CDR3 derived from a VL having the amino acid sequence of SEQ ID NO: 21, and the VH comprises VH CDR1, CDR2 and CDR3 derived from a VH having the amino acid sequence of SEQ ID NO: 22. In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to MASP-2 is provided, comprising a VL and a VH, wherein the VL comprises VL CDR1, CDR2 and CDR3 derived from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2 and CDR3 derived from a VH having the amino acid sequence of SEQ ID NO: 24. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 25. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 27. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 28. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 29.In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 30. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 31. In some embodiments, the VL comprises VL CDR1, CDR2, and CDR3 from a VL having the amino acid sequence of SEQ ID NO: 23, and the VH comprises VH CDR1, CDR2, and CDR3 from a VH having the amino acid sequence of SEQ ID NO: 32.

[0129]

[0135] In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein is an antibody designated as 3E10. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein has a VL derived from 3E10 (SEQ ID NO: 21). In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein has a VH derived from 3E10 (SEQ ID NO: 22). The anti-MASP-2 antibody or antigen-binding fragment thereof provided herein may have both a VL and a VH derived from 3E10. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein has a VL comprising VL CDRs 1, 2, and 3 derived from the VL derived from 3E10 (SEQ ID NO: 21). In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein has a VH comprising VH CDRs 1, 2, and 3 derived from the VH derived from 3E10 (SEQ ID NO: 22). The anti-MASP-2 antibodies or antigen-binding fragments thereof provided herein may have a VL comprising VL CDRs 1, 2, and 3 derived from the VL of 3E10, and a VH comprising VH CDRs 1, 2, and 3 derived from the VH of 3E10. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments thereof provided herein are variants of 3E10. The variants of 3E10 may have a VL that is a variant of the VL of 3E10 with up to about five amino acid substitutions, additions, and / or deletions in SEQ ID NO: 21. The variants of 3E10 may have a VH that is a variant of the VH of 3E10 with up to about five amino acid substitutions, additions, and / or deletions in SEQ ID NO: 22. The amino acid substitutions, additions, and / or deletions may be within the VH CDR or the VL CDR. In some embodiments, the amino acid substitutions, additions, and / or deletions are not present in the CDRs. In some embodiments, the 3E10 variant has up to about 5 conservative amino acid substitutions. In some embodiments, the 3E10 variant has up to 3 conservative amino acid substitutions. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein is a humanized antibody or antigen-binding fragment derived from 3E10.In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein is a human antibody or antigen-binding fragment derived from 3E10.

[0130]

[0136] In some embodiments, a humanized 3E10 is provided. In some embodiments, the humanized anti-MASP-2 antibody or antigen-binding fragment thereof provided herein comprises a VL having the amino acid sequence of SEQ ID NO: 23. In some embodiments, the humanized anti-MASP-2 antibody or antigen-binding fragment thereof provided herein comprises a VH having an amino acid sequence selected from SEQ ID NOs: 24 to 32. In some embodiments, the humanized anti-MASP-2 antibody or antigen-binding fragment thereof provided herein comprises a VL having the amino acid sequence of SEQ ID NO: 23 and a VH having an amino acid sequence selected from SEQ ID NOs: 24 to 32. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment thereof provided herein is a variant of the humanized 3E10 provided herein. The variant may have a VL that is a variant of the VL of humanized 3E10, having up to about five amino acid substitutions, additions, and / or deletions in the amino acid sequence of SEQ ID NO: 23. The variant may have a VH that is a variant of the VH of humanized 3E10 having up to about 5 amino acid substitutions, additions, and / or deletions in an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 32. In some embodiments, the humanized 3E10 variant has up to about 5 conservative amino acid substitutions.

[0131]

[0137] In some embodiments, antibodies or antigen-binding fragments that compete with the above-mentioned antibodies or antigen-binding fragments for binding to MASP-2 (e.g., human MASP-2) are also provided. An antibody that "competes with another antibody for binding to a target" refers to an antibody that inhibits (partially or completely) the binding of the other antibody to a target. Whether two antibodies compete with each other for binding to a target, i.e., whether and to what extent one antibody inhibits the binding of the other antibody to a target, can be determined using known competition experiments, such as BIACORE® surface plasmon resonance (SPR) analysis. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment competes with another antibody or antigen-binding fragment for binding to MASP-2 and inhibits the binding of the other antibody or antigen-binding fragment to MASP-2 by at least 50%, 60%, 70%, 80%, 90%, or 100%. Competitive assays can be performed, for example, as described in Cold Spring Haib Protoc; 2006 by Ed Harlow and David Lane; doi: 10.1101 / pdb.prot 4277, or Chapter 11 of Using Antibodies, by Ed Harlow and David Lane, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA 1999.

[0132]

[0138] In some embodiments, an antibody or antigen-binding fragment is provided that competes with the anti-MASP-2 antibody or antigen-binding fragment disclosed herein for binding to human MASP-2. In some embodiments, an antibody or antigen-binding fragment is provided that competes with 3E10 for binding to human MASP-2. In some embodiments, an antibody or antigen-binding fragment is provided that competes with humanized 3E10 disclosed herein for binding to human MASP-2.

[0133]

[0139] The present disclosure further contemplates additional variants and equivalents that are substantially homologous to the recombinant, monoclonal, chimeric, humanized, and human antibodies described herein, or antibody fragments thereof. In some embodiments, it is desirable to improve the binding affinity of the antibody. In some embodiments, it is desirable to modulate the biological properties of the antibody, including, but not limited to, specificity, thermal stability, expression level, effector function, glycosylation, immunogenicity, and / or solubility. Those skilled in the art will understand that amino acid changes can alter post-translational processing of the antibody, for example, by changing the number or location of glycosylation sites or altering membrane anchoring characteristics.

[0134]

[0140] Mutations can be substitutions, deletions, or insertions of one or more nucleotides encoding the antibody or polypeptide, resulting in a change in the amino acid sequence when compared to the native antibody or polypeptide sequence. In some embodiments, amino acid substitutions result in the replacement of one amino acid with another amino acid having similar structural and / or chemical properties, e.g., conservative amino acid substitutions, such as a leucine for a serine. Insertions or deletions can range from about 1 to 5 amino acids. In some embodiments, substitutions, deletions, or insertions comprise fewer than 25 amino acid substitutions, fewer than 20 amino acid substitutions, fewer than 15 amino acid substitutions, fewer than 10 amino acid substitutions, fewer than 5 amino acid substitutions, fewer than 4 amino acid substitutions, fewer than 3 amino acid substitutions, or fewer than 2 amino acid substitutions compared to the parent molecule. In some embodiments, biologically useful and / or relevant amino acid sequence variations can be determined by systematically making insertions, deletions, or substitutions in the sequence and testing the resulting mutant proteins for activity compared to the parent protein.

[0135]

[0141] It is known in the art that the constant region(s) of an antibody mediate several effector functions, and these effector functions may vary depending on the antibody isotype. For example, binding of the C1 component of complement to the Fc region of an IgG or IgM antibody (bound to an antigen) activates the complement system. Complement activation is important in the opsonization and lysis of cellular pathogens. Complement activation also stimulates the inflammatory response and may be involved in autoimmune hypersensitivity. Furthermore, the Fc region of an antibody can bind to cells expressing Fc receptors (FcRs). There are several Fc receptors specific for different classes of antibodies, including IgG (gamma receptors), IgE (epsilon receptors), IgA (alpha receptors), and IgM (mu receptors). Binding of antibodies to Fc receptors on cell surfaces elicits several important and diverse biological responses, including the uptake and destruction of antibody-coated particles, clearance of immune complexes, lysis of antibody-coated target cells by killer cells (called antibody-dependent cellular cytotoxicity or ADCC), release of inflammatory mediators, placental transfer, and regulation of immunoglobulin production.

[0136]

[0142] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgA antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgD antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgE antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgG antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgM antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgG1 antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgG2 antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgG3 antibody. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise at least one constant region of a human IgG4 antibody.

[0137]

[0143] In some embodiments, at least one or more of the constant regions in the anti-MASP-2 antibodies or antigen-binding fragments described herein have been modified or deleted. In some embodiments, the antibodies include modifications to one or more of the three heavy chain constant regions (CH1, CH2, or CH3) and / or the light chain constant region (CL). In some embodiments, the heavy chain constant region of the modified antibody includes at least one human constant region. In some embodiments, the heavy chain constant region of the modified antibody includes more than one human constant region. In some embodiments, modifications to the constant region include the addition, deletion, or substitution of one or more amino acids in one or more regions. In some embodiments, one or more regions are partially or entirely deleted from the constant region of the modified antibody. In some embodiments, the entire CH2 domain has been removed from the antibody (ΔCH2 construct). In some embodiments, the deleted constant region is replaced by a short amino acid spacer that provides some of the molecular flexibility normally conferred by the deleted constant region. In some embodiments, the modified antibody includes a CH3 domain fused directly to the hinge region of the antibody. In some embodiments, the modified antibody comprises a peptide spacer inserted between the hinge region and the modified CH2 and / or CH3 domain.

[0138]

[0144] In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment comprises an Fc region. In some embodiments, the Fc region is fused via a hinge. The hinge may be an IgG1 hinge, an IgG2 hinge, or an IgG3 hinge. The amino acid sequences of the Fc regions of human IgG1, IgG2, IgG3, and IgG4 are known to those skilled in the art. In some cases, Fc regions with amino acid mutations have been identified in native antibodies. In some embodiments, a modified antibody (e.g., a modified Fc region) provides altered effector function, which in turn affects the biological profile of the antibody. For example, in some embodiments, deletion or inactivation of the constant region (by point mutation or other means) reduces Fc receptor binding when the modified antibody circulates. In some embodiments, the constant region modification reduces the immunogenicity of the antibody. In some embodiments, the constant region modification increases the serum half-life of the antibody. In some embodiments, the constant region modification reduces the serum half-life of the antibody. In some embodiments, the constant region modifications reduce or eliminate ADCC and / or complement-dependent cytotoxicity (CDC) of the antibody. In some embodiments, specific amino acid substitutions in the human IgG1 Fc region with corresponding IgG2 or IgG4 residues reduce effector function (e.g., ADCC and CDC) in the modified antibody. In some embodiments, the antibody does not have one or more effector functions (e.g., an "effector-less" antibody). In some embodiments, the antibody does not have ADCC activity and / or CDC activity. In some embodiments, the antibody does not bind to Fc receptors and / or complement factors. In some embodiments, the antibody does not have effector function(s). In some embodiments, the constant region modifications increase or enhance ADCC and / or CDC of the antibody. In some embodiments, the constant region is modified to eliminate disulfide bonds or oligosaccharide moieties. In some embodiments, the constant region is modified to add / substitute one or more amino acids to provide one or more cytotoxin, oligosaccharide, or carbohydrate attachment sites.In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment comprises a variant Fc region that is modified with substitutions at specific amino acid positions compared to the native Fc region. In some embodiments, the anti-MASP-2 antibody or antigen-binding fragment described herein comprises an IgG1 heavy chain constant region containing one or more amino acid substitutions selected from the group consisting of K214, L234, L235, G237, D356, and L358, numbered according to the EU index. The K214 substitution may be, for example, K214R. The L234 substitution may be, for example, L234A or L234E. The L235 substitution may be, for example, L235A or L235E. The G237 substitution may be, for example, G237A. The D356 substitution may be, for example, D356E. The L358 substitution may be, for example, L358M. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise an IgG1 heavy chain constant region comprising one or more amino acid substitutions selected from the group consisting of K214R, L234A, L235E, G237A, D356E, and L358M, numbered according to the EU index. In some embodiments, the IgG1 heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of K214R, L234A, L235E, G237A, A330S, P331S, D356E, and L358M, numbered according to the EU index. In some embodiments, the IgG1 heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of K214R, C226S, C229S, and P238S, numbered according to the EU index. In some embodiments, the IgG1 heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of K214R, D356E, and L358M, numbered according to the EU index. In some embodiments, the IgG1 heavy chain constant region comprises one or more amino acid substitutions selected from the group consisting of S131C, K133R, G137E, G138S, Q196K, I199T, N203D, K214R, C226S, C229S, and P238S, numbered according to the EU index.

[0139]

[0145] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein comprise an IgG4 heavy chain constant region containing one or more amino acid substitutions that extend the terminal half-life of the antibody or antigen-binding fragment. As understood in the art, terminal half-life, also known as biological half-life, refers to the time required for plasma / blood concentration to decrease by 50% after administration and after reaching pseudo-equilibrium of distribution. The one or more amino acid substitutions may be selected from the group consisting of S228, F234, L235, M252, S254, T256, K288, T307, M428, N434, H435, and Y436, numbered according to the EU index. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at S228. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at F234. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at L235. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at M252. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at S254. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at T256. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at K288. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at T307. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at M428. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at N434. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at H435. In some embodiments, the IgG4 heavy chain constant region has an amino acid substitution at Y436. The amino acid substitution at S228 can be, for example, S228P. The amino acid substitution at F234 can be, for example, F234A. The amino acid substitution at L235 can be, for example, L235A. The amino acid substitution at M252 can be, for example, M252Y. The amino acid substitution at S254 can be, for example, S254T. The amino acid substitution at T256 can be, for example, T256E. The amino acid substitution at K288 can be, for example, K288A.The amino acid substitution at T307 can be, for example, T307Q or T307H. The amino acid substitution at M428 can be, for example, M428L. The amino acid substitution at N434 can be, for example, N434A. The amino acid substitution at H435 can be, for example, H435A or H435R. The amino acid substitution at Y436 can be, for example, Y436T or Y436F.

[0140]

[0146] In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise: i) S228P; ii) F234A and L235A; iii) S228P, F234A and L235A; (iv) T307H and N434A; v) M252Y, S254T and T256E; vi) M428L, N434A and Y436T; viii) S228P, M2 viii) S228P, F234A, L235A, M252Y, S254T and T256E; viii) S228P, F234A, L235A, M252Y, S254T and T256E; ix) S228P, F234A, L235A, T307Q and N434A; and (x) M252Y, S254T, T307H and N434A.

[0141]

[0147] In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with the amino acid substitution S228P. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with the amino acid substitutions F234A and L235A. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with the amino acid substitutions S228P, F234A, and L235A. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with the amino acid substitutions T307H and N434A. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with the amino acid substitutions M252Y, S254T, and T256E. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with amino acid substitutions M428L, N434A, and Y436T. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with amino acid substitutions S228P, M252Y, S254T, and T256E. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with amino acid substitutions S228P, F234A, L235A, M252Y, S254T, and T256E. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with amino acid substitutions S228P, F234A, L235A, T307Q, and N434A. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein comprise a mutant human IgG4 heavy chain modified with amino acid substitutions M252Y, S254T, T307H and N434A.

[0142]

[0148] In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 33, or a variant thereof having an amino acid sequence that is at least about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identical to SEQ ID NO: 33.

[0143]

[0149] In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 34, or a variant thereof having an amino acid sequence at least about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identical to SEQ ID NO: 34. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having an amino acid sequence selected from the group consisting of SEQ ID NOs: 34-74. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 34. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 35. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 36. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 37. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 38. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 39. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 40. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 41. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 42. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 43. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 44. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 45.In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 46. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 47. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 48. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 49. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 50. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 51. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 52. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 53. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 54. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 55. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 56. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 57. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 58. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 59. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 60. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 61.In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 62. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 63. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 64. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 65. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 66. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 67. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 68. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 69. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 70. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 71. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 72. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 73. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a heavy chain having the amino acid sequence of SEQ ID NO: 74.

[0144]

[0150] In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain comprising the amino acid sequence of SEQ ID NO: 33, or a variant thereof having an amino acid sequence that is at least about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identical to SEQ ID NO: 33, and a heavy chain comprising the amino acid sequence of SEQ ID NO: 34, or a variant thereof having an amino acid sequence that is at least about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or 100% identical to SEQ ID NO: 34.

[0145]

[0151] In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 34. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 35. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 36. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 37. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 38. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 39. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 40. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 41. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 42. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 43. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 44. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 45.In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 46. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 47. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 48. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 49. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 50. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 51. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 52. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 53. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 54. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 55. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 56. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 57.In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 58. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 59. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 60. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 61. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 62. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 63. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 64. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 65. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 66. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 67. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 68. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 69.In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 70. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 71. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 72. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 73. In some embodiments, an anti-MASP-2 antibody or antigen-binding fragment is provided, comprising a light chain having the amino acid sequence of SEQ ID NO: 33 and a heavy chain having the amino acid sequence of SEQ ID NO: 74.

[0146]

[0152] In some embodiments, variants may include the addition of amino acid residues at the amino and / or carboxyl termini of the antibody or polypeptide. The length of the additional amino acid residues may range from 1 to 100 or more residues. In some embodiments, variants include an N-terminal methionyl residue. In some embodiments, variants include an additional polypeptide / protein (e.g., an Fc region) to generate a fusion protein. In some embodiments, variants may be modified to be detectable and include a detectable label and / or protein (e.g., a fluorescent tag or an enzyme).

[0147]

[0153] The variant antibodies or antigen-binding fragments described herein can be generated using methods known in the art, including but not limited to site-directed mutagenesis, alanine scanning mutagenesis, and PCR mutagenesis.

[0148]

[0154] In some embodiments, variants of anti-MASP-2 antibodies or antigen-binding fragments disclosed herein may retain the ability to bind to MASP-2 to a similar, the same, or greater extent than the parent antibody or antigen-binding fragment. In some embodiments, the amino acid sequence of the variant may be at least about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more identical to that of the parent antibody or antigen-binding fragment. In certain embodiments, variants of anti-MASP-2 antibodies or antigen-binding fragments comprise the amino acid sequence of the parent anti-MASP-2 antibody or antigen-binding fragment with one or more conservative amino acid substitutions. Conservative amino acid substitutions are known in the art and include amino acid substitutions in which an amino acid having particular physical and / or chemical properties is replaced with another amino acid having the same or similar physical or chemical properties.

[0149]

[0155] In some embodiments, a variant of an anti-MASP-2 antibody or antigen-binding fragment comprises the amino acid sequence of a parent antibody or antigen-binding fragment with one or more non-conservative amino acid substitutions. In some embodiments, a variant of an anti-MASP-2 antibody or antigen-binding fragment comprises the amino acid sequence of a parent antibody or antigen-binding fragment with one or more non-conservative amino acid substitutions, wherein the one or more non-conservative amino acid substitutions do not interfere with or inhibit one or more biological activities of the variant (e.g., MASP-2 binding). In certain embodiments, the one or more conservative amino acid substitutions and / or one or more non-conservative amino acid substitutions can enhance the biological activity of the variant such that the biological activity of the functional variant is improved compared to the parent antibody or antigen-binding fragment.

[0150]

[0156] In some embodiments, the variants may have one, two, three, four, or five amino acid substitutions in the CDRs of the binding moiety (e.g., VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3).

[0151]

[0157] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein are chemically modified, either naturally or by intervention. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments are chemically modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, and / or linkage to a cellular ligand or other protein. Any of a number of chemical modifications can be performed by known techniques. The anti-MASP-2 antibodies or antigen-binding fragments may include one or more analogs of amino acids (including, for example, unnatural amino acids), as well as other modifications known in the art.

[0152]

[0158] Epitope mapping is a method for identifying the binding site, region, or epitope on a target protein to which an antibody binds. Various methods for mapping epitopes on a target protein are known in the art. These methods include mutagenesis, including but not limited to shotgun mutagenesis, site-directed mutagenesis, and alanine scanning; domain or fragment scanning; peptide scanning (e.g., Pepscan technology); display methods (e.g., phage display, microbial display, and ribosome / mRNA display); methods involving proteolysis and mass spectrometry; and structure determination (e.g., X-ray crystallography and NMR). In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein are characterized by assays including, but not limited to, N-terminal sequencing, amino acid analysis, HPLC, mass spectrometry, ion-exchange chromatography, and papain digestion.

[0153]

[0159] As described in further detail in the experiments below, antibody 3E10 and humanized 3E10 provided herein bind to the protease domain of human MASP-2 (amino acid residues 415 to 671 of SEQ ID NO: 1) and do not compete with the benchmark antibody narsoplimab for binding to human MASP-2. In some embodiments, anti-MASP-2 antibodies and antigen-binding fragments are provided that bind to the protease domain of human MASP-2. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments provided herein do not detectably bind to truncated human MASP-2 lacking the protease domain. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments provided herein do not compete with narsoplimab for binding to human MASP-2.

[0154]

[0160] Anti-MASP-2 antibodies or antigen-binding fragments of the present disclosure can be analyzed for their physical, chemical, and / or biological properties by various methods known in the art. In some embodiments, anti-MASP-2 antibodies are tested for their ability to bind to MASP-2 (e.g., human MASP-2). Binding assays include, but are not limited to, BLI, SPR (e.g., Biacore), ELISA, and FACS. Additionally, antibodies can be evaluated for solubility, stability, thermal stability, viscosity, expression level, expression quality, and / or purification efficiency.

[0155]

[0161] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein bind with high affinity, e.g., at least 10 -7 M or less, 10 -8 M or less, 5×10 -9 M or less, 10 -9 M or less, 5×10 -10 M or less, 10 -10 M or less, 5×10 -11 M or less, 10 -11 M or less, 5×10 -12 M or less, 10 -12 M or less, 10 -12 M~10 -7 M, 10-11 M~10 -7 M, 10 -10 M~10 -7 M, 10 -9 M~10 -7 M, 10 -8 M~10 -7 M, 10 -10 M~10 -8 M, 10 -9 M~10 -8 M, 10 -11 M~10 -9 M or 10 -10 M~10 -9 K of M D In some embodiments, K D is determined by BLI. In some embodiments, K D is determined by SPR. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein have high affinity, e.g., 10 as measured by SPR. -7 M or less, 10 -8 M or less, 5×10 -9 M or less, 10 -9 M or less, 5×10 -10 M or less, 10 -10 M or less, 5×10 -11 M or less, 10 -11 M or less, 5×10 -12 M or less, 10 -12 M or less, 10 -12 M~10 -7 M, 10 -11 M~10 -7 M, 10 -10 M~10 -7 M, 10 -9 M~10 -7 M, 10 -8 M~10 -7 , 10 -10 M~10 -8 M, 10 -9 M~10 -8 M, 10 -11 M~10 -9 M or 10 -10 M~10 -9 K of M D It binds to human MASP-2.

[0156]

[0162] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein have a binding activity of 10 -9 K below M D In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein bind to human MASP-2 at 10 as measured by SPR. -10 K below M D In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein bind to human MASP-2 at 10 as measured by SPR. -11 M~10 -09 K is the range of M D In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein bind to human MASP-2 at a concentration of 5×10 as measured by SPR. -11 M~10 -09 K is the range of M D In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein bind to human MASP-2 at a concentration of 5×10 as measured by SPR. -11 M~5×10 -10 K is the range of M D In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein bind to human MASP-2 at a binding affinity of about 10 as measured by SPR. -10 K of M D It binds to human MASP-2.

[0157]

[0163] The anti-MASP-2 antibodies and antigen-binding fragments described herein are capable of inhibiting MASP-2-dependent complement activation. In other words, the anti-MASP-2 antibodies and antigen-binding fragments described herein are capable of inhibiting the lectin pathway of complement activation. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments provided herein are capable of inhibiting MASP-2-dependent complement activation by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90%. In one embodiment, the anti-MASP-2 antibodies or antigen-binding fragments provided herein reduce MASP-2-dependent complement activation by at least 50% (i.e., MASP-2-dependent complement activation is 50% or less compared to the activation level in the absence of the antibody or antigen-binding fragment). In one embodiment, the anti-MASP-2 antibody or antigen-binding fragment provided herein reduces MASP-2-dependent complement activation by at least 80% (i.e., MASP-2-dependent complement activation is 20% or less compared to the activation level in the absence of the antibody or antigen-binding fragment). In one embodiment, the anti-MASP-2 antibody or antigen-binding fragment provided herein reduces MASP-2-dependent complement activation by at least 90% (i.e., MASP-2-dependent complement activation is 10% or less compared to the activation level in the absence of the antibody or antigen-binding fragment).

[0158]

[0164] In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein do not inhibit the classical or alternative pathways of complement activation.

[0165] The inhibitory effect of the anti-MASP-2 antibodies and antigen-binding fragments provided herein on MASP-2-dependent complement activation can be measured at multiple levels, such as C4 activation, C3 activation, and MAC activation. For example, various forms of assays for measuring C4 activation, C3 activation, and MAC activation are known in the art (see, for example, WO2012 / 151481), and some assays are described in the Examples below.

[0159]

[0166] The anti-MASP-2 antibodies and antigen-binding fragments described herein can inhibit C4 activation. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, or 0.001 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of about 0.2 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of about 0.1 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of about 0.05 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of about 0.01 μg / mL or less, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001-0.5 μg / mL, 0.001-0.2 μg / mL, 0.001-0.1 μg / mL, 0.001-0.05 μg / mL, 0.001-0.01 μg / mL, 0.001-0.005 μg / mL, 0.005-0.5 μg / mL, 0.005-0.2 μg / mL, 0.005-0.1 μg / mL, 0.005-0.05 μg / mL, 0.005-0.01 μg / mL, or 0.001-0.5 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001 to 0.1 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001 to 0.01 μg / mL, as measured, for example, in an in vitro assay. 50 can inhibit the activation of C4.

[0160]

[0167] C4 activation assays can be performed in the presence of various concentrations of human serum (e.g., 1-100% or 5-50%) to measure the inhibitory effect of anti-MASP-2 antibodies or antigen-binding fragments. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, or 0.001 μg / mL or less in the presence of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, or 0.001 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, or 0.001 μg / mL or less in the presence of about 10% human serum, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, or 0.001 μg / mL or less, as measured, for example, in an in vitro assay. 50 can inhibit the activation of C4.

[0161]

[0168] In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.2 μg / mL or less in the presence of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.1 μg / mL or less in the presence of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.05 μg / mL or less in the presence of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. 50 can inhibit the activation of C4.

[0162]

[0169] In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein exhibit a concentration of 0.001 to 0.5 μg / mL, 0.001 to 100% human serum, specifically, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. IC in the range of 0.2μg / mL, 0.001-0.1μg / mL, 0.001-0.05μg / mL, 0.001-0.01μg / mL, 0.001-0.005μg / mL, 0.005-0.5μg / mL, 0.005-0.2μg / mL, 0.005-0.1μg / mL, 0.005-0.05μg / mL, 0.005-0.01μg / mL, or 0.001-0.5μg / mL 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001 to 0.1 μg / mL in the presence of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.01 to 0.1 μg / mL in the presence of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% human serum, as measured, for example, in an in vitro assay. 50 can inhibit the activation of C4.

[0163]

[0170] In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.001 to 0.1 μg / mL in the presence of about 5% human serum, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.01 to 0.1 μg / mL in the presence of about 5% human serum, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001 to 0.1 μg / mL in the presence of about 10% human serum, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.01 to 0.1 μg / mL in the presence of about 10% human serum, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.001 to 0.1 μg / mL in the presence of about 50% human serum, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.01 to 0.1 μg / mL in the presence of about 50% human serum, as measured, for example, in an in vitro assay. 50 can inhibit the activation of C4.

[0164]

[0171] The anti-MASP-2 antibodies and antigen-binding fragments described herein can inhibit C3 activation. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, 0.002 μg / mL or less, or 0.001 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.2 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.1 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.05 μg / mL or less, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001-0.5 μg / mL, 0.001-0.2 μg / mL, 0.001-0.1 μg / mL, 0.001-0.05 μg / mL, 0.001-0.01 μg / mL, 0.001-0.005 μg / mL, 0.005-0.5 μg / mL, 0.005-0.2 μg / mL, 0.005-0.1 μg / mL, 0.005-0.05 μg / mL, 0.005-0.01 μg / mL, or 0.001-0.5 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001 to 0.1 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001 to 0.05 μg / mL, as measured, for example, in an in vitro assay. 50 can inhibit the activation of C3.

[0165]

[0172] The anti-MASP-2 antibodies and antigen-binding fragments described herein can inhibit MAC activation. In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.5 μg / mL or less, 0.4 μg / mL or less, 0.3 μg / mL or less, 0.2 μg / mL or less, 0.1 μg / mL or less, 0.05 μg / mL or less, 0.04 μg / mL or less, 0.03 μg / mL or less, 0.02 μg / mL or less, 0.01 μg / mL or less, 0.005 μg / mL or less, 0.002 μg / mL or less, or 0.001 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.2 μg / mL or less, as measured, for example, in an in vitro assay. 50In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.1 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC of 0.05 μg / mL or less, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 of 0.001-0.5 μg / mL, 0.001-0.2 μg / mL, 0.001-0.1 μg / mL, 0.001-0.05 μg / mL, 0.001-0.01 μg / mL, 0.001-0.005 μg / mL, 0.005-0.5 μg / mL, 0.005-0.2 μg / mL, 0.005-0.1 μg / mL, 0.005-0.05 μg / mL, 0.005-0.01 μg / mL, or 0.001-0.5 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 activity in the range of 0.001 to 0.1 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 activity in the range of 0.001 to 0.05 μg / mL, as measured, for example, in an in vitro assay. 50 In some embodiments, the anti-MASP-2 antibodies and antigen-binding fragments described herein have an IC50 activity in the range of 0.001 to 0.01 μg / mL, as measured, for example, in an in vitro assay. 50 can inhibit the activation of MAC.

[0166]

[0173] Thus, in some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments thereof described herein have (1) a K of about 1 nM or less as measured by SPR.D (2) binds to human MASP-2 with an IC of approximately 0.1 μg / mL or less measured in vitro 50 (3) an IC of approximately 0.1 μg / mL or less measured in vitro; 50 (4) an IC of approximately 0.1 μg / mL or less measured in vitro; 50 (5) inhibiting MAC activation; (6) not affecting the classical or alternative pathways of complement activation; or (7) having any combination of the properties of (1) to (5).

[0167]

[0174] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments thereof described herein have (1) a K of about 1 nM or less as measured by SPR. D and (2) an IC of about 0.1 μg / mL or less as measured in vitro. 50 can inhibit the activation of C4.

[0168]

[0175] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to the protease domain of human MASP-2 is provided, the antibody or antigen-binding fragment thereof comprising: (1) K measured by SPR of approximately 1 nM or less D binds to human MASP-2 (2) IC measured in vitro of approximately 0.1 μg / mL or less 50 inhibits the activation of C4, (3) IC measured in vitro of approximately 0.1 μg / mL or less 50 inhibits C3 activation, (4) IC measured in vitro of approximately 0.1 μg / mL or less 50 inhibits MAC activation, (5) does not affect the classical or alternative pathways of complement activation; or (6) Possessing any combination of the properties of (1) to (5).

[0169]

[0176] In some embodiments, an antibody or antigen-binding fragment thereof that specifically binds to the protease domain of human MASP-2, and has a K of 1 nM or less as measured by SPR. D In some embodiments, the present invention provides an antibody or antigen-binding fragment thereof that specifically binds to the protease domain of human MASP-2 and has an IC of 0.1 μg / mL or less as measured in vitro. 50 In some embodiments, the present invention provides an antibody or antigen-binding fragment thereof that specifically binds to the protease domain of human MASP-2 and has an IC of 0.1 μg / mL or less as measured in vitro. 50 In some embodiments, the present invention provides an antibody or antigen-binding fragment thereof that specifically binds to the protease domain of human MASP-2 and has an IC of 0.1 μg / mL or less as measured in vitro. 50 In some embodiments, the antibody or antigen-binding fragment thereof does not affect the classical or alternative pathway of complement activation.

[0170]

[0177] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein are conjugated to a detectable substance or molecule, allowing the agent to be used for diagnosis and / or detection. Detectable substances include enzymes, such as horseradish peroxidase, alkaline phosphatase, β-galactosidase, and acetylcholinesterase; prosthetic groups, such as biotin and flavins; fluorescent substances, such as umbelliferone, fluorescein, fluorescein isothiocyanate (FITC), rhodamine, tetramethylrhodamine isothiocyanate (TRITC), dichlorotriazinylamine fluorescein, dansyl chloride, cyanine (Cy3), and phycoerythrin; bioluminescent substances, such as luciferase; and radioactive substances, such as 212 Bi, 14 C. 57 Co,51 Cr, 67 Cu, 18 F, 68 Ga, 67 Ga, 153 Gd, 159 Gd, 68 Ge, 3 H, 166 Ho, 131 I, 125 I, 123 I, 121 I, 115 In, 113 In, 112 In, 111 In, 140 La, 177 Lu, 54 Mn, 99 Mo, 32 P, 103 Pd, 149 Pm, 142 Pr, 186 Re, 188 Re, 105 Rh, 97 Ru, 35 S, 47 Sc, 75 Se, 153 Sm, 113 Sn, 117 Sn, 85 Sr, 99m Tc, 201 Ti, 133 Xe, 90 Y, 69 Yb, 175 Yb, 65 Zn; positron emitting metals; and magnetic metal ions. Positron emitting metals; and magnetic metal ions.

[0171]

[0178] The anti-MASP-2 antibodies or antigen-binding fragments described herein may be bound to a solid support. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride, or polypropylene. In some embodiments, the immobilized anti-MASP-2 antibodies or antigen-binding fragments are used in immunoassays. In some embodiments, the immobilized anti-MASP-2 antibodies or antigen-binding fragments are used in the purification of target antigens (e.g., human MASP-2). 5.4 Polynucleotides and Vectors

[0179] Also provided herein are polynucleotides encoding the polypeptides described herein (e.g., anti-MASP-2 antibodies or antigen-binding fragments). The term "polynucleotide encoding a polypeptide" encompasses polynucleotides that contain only the coding sequence for the polypeptide, as well as polynucleotides that contain additional coding and / or non-coding sequences. Polynucleotides of the present disclosure may be in the form of RNA or in the form of DNA. DNA may be cDNA, genomic DNA, or synthetic DNA, and may be double-stranded or single-stranded. Single-stranded DNA may be the coding strand or the non-coding (antisense) strand. Polynucleotides of the present disclosure may be mRNA.

[0172]

[0180] Expressly contemplated herein are polynucleotides encoding any anti-MASP-2 antibody or antigen-binding fragment disclosed herein. By way of illustration, in some embodiments, the polynucleotides provided herein encode anti-MASP-2 antibodies or antigen-binding fragments, which comprise: (1) a light chain variable region (VL) comprising, as defined by Kabat, (a) a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9, and 11, respectively, or variants thereof having up to about five amino acid substitutions, additions, and / or deletions in the VL CDRs; and / or (b) a heavy chain variable region (VH) comprising, as defined by IMGT, (a) a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 12, 16, and 18, respectively, or variants thereof having up to about five amino acid substitutions, additions, and / or deletions in the VH CDRs; or (2) a heavy chain variable region (VH) comprising, as defined by IMGT, (a) a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 8, 10, and 11, respectively, or variants thereof having up to about five amino acid substitutions, additions, and / or deletions in the VH CDRs. and / or (b) a VL comprising a VH CDR1, a VH CDR2 and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 13, 17 and 19, respectively, or variants thereof having up to about 5 amino acid substitutions, additions and / or deletions in the VH CDRs.

[0173]

[0181] In some embodiments, the polynucleotides provided herein encode an anti-MASP-2 antibody or antigen-binding fragment comprising (a) a VL having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 23, and / or (b) a VH having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 24. The polynucleotide may be in the form of DNA. The polynucleotide may be in the form of mRNA.

[0174]

[0182] In some embodiments, the polynucleotides provided herein encode an anti-MASP-2 antibody or antigen-binding fragment disclosed herein comprising a VL and a VH, wherein the VL comprises a VL CDR1, CDR2, and CDR3, and the VH comprises a VH CDR1, CDR2, and CDR3, and wherein the VL CDR1, VL CDR2, VL CDR3, VH CDR1, VH CDR2, and VH CDR3 has the amino acid sequence of (1) SEQ ID NO: 7, 9, 11, 12, 16, and 18, respectively; (2) SEQ ID NO: 8, 10, 11, 13, 17, and 19, respectively; (3) SEQ ID NO: 8, 10, 11, 14, 17, and 19, respectively; (4) SEQ ID NO: 8, 10, 11, 15, 17, and 19, respectively; (5) SEQ ID NO: 8, 10, 11, 13, 17, and 20, respectively; (6) SEQ ID NO: 8, 10, 11, 14, 17, and 20, respectively; (7) SEQ ID NO: 8, 10, 11, 15, 17, and 20, respectively, or a variant thereof having up to about five amino acid substitutions, additions, and / or deletions in the CDR. The polynucleotide may be in the form of DNA. The polynucleotide may be in the form of mRNA.

[0175]

[0183] In some embodiments, the polynucleotide provided herein encodes an anti-MASP-2 antibody or antigen-binding fragment disclosed herein comprising a VL and a VH, wherein the VL and VH have the amino acid sequences of (1) SEQ ID NOs: 21 and 22, respectively, (2) SEQ ID NOs: 23 and 24, respectively, (3) SEQ ID NOs: 23 and 25, respectively, (4) SEQ ID NOs: 23 and 26, respectively, (5) SEQ ID NOs: 23 and 27, respectively, (6) SEQ ID NOs: 23 and 28, respectively, (7) SEQ ID NOs: 23 and 29, respectively, (8) SEQ ID NOs: 23 and 30, respectively, (9) SEQ ID NOs: 23 and 31, respectively, or (10) SEQ ID NOs: 23 and 32, respectively. The polynucleotide may be in the form of DNA. The polynucleotide may be in the form of mRNA.

[0176]

[0184] In some embodiments, the VL and VH are linked by a linker. The linker can be a flexible linker or a rigid linker. In some embodiments, the linker has the amino acid sequence of (GGGGS)n, where n=1, 2, 3, 4, or 5 (SEQ ID NO: 77). In some embodiments, the linker has the amino acid sequence of (EAAAK)n, where n=1, 2, 3, 4, or 5 (SEQ ID NO: 78). In some embodiments, the linker has the amino acid sequence of (PA)nP, where n=1, 2, 3, 4, or 5 (SEQ ID NO: 79).

[0177]

[0185] The present disclosure also provides variants of the polynucleotides described herein, which, for example, encode fragments, analogs, and / or derivatives of the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein. In some embodiments, the present disclosure provides polynucleotides having a nucleotide sequence that is at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, or at least about 99% identical to the polynucleotide sequence encoding the anti-MASP-2 antibodies or antigen-binding fragments described herein. In some embodiments, the present disclosure provides polynucleotides having a nucleotide sequence that is at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, or at least about 99% identical to the polynucleotide sequence encoding the anti-MASP-2 antibodies or antigen-binding fragments described herein.

[0178]

[0186] As used herein, the phrase "a polynucleotide having a nucleotide sequence at least about 95% identical to a polynucleotide sequence" means that the nucleotide sequence of the polynucleotide is identical to the reference sequence, except that the polynucleotide sequence may contain up to 5 point mutations per 100 nucleotides of the reference nucleotide sequence. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence can be deleted or replaced with other nucleotides, or up to 5% of the total number of nucleotides in the reference sequence can be inserted into the reference sequence. These mutations of the reference sequence can occur at the 5' or 3' terminal position of the reference nucleotide sequence, or at any position between these terminal positions, and can be individually interspersed among the nucleotides in the reference sequence or interspersed within one or more consecutive groups within the reference sequence.

[0179]

[0187] Polynucleotide variants may contain alterations in coding regions, non-coding regions, or both. In some embodiments, polynucleotide variants contain alterations that produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide. In some embodiments, polynucleotide variants contain silent substitutions that do not result in changes to the amino acid sequence of a polypeptide (due to the degeneracy of the genetic code). Polynucleotide variants can be produced for a variety of reasons, such as to optimize codon expression for a particular host (e.g., changing codons in human mRNA to codons preferred by a bacterial host such as E. coli). In some embodiments, polynucleotide variants contain at least one silent mutation in a non-coding or coding region of the sequence.

[0180]

[0188] In some embodiments, polynucleotide variants are generated to modify or alter the expression (or expression levels) of the encoded polypeptide. In some embodiments, polynucleotide variants are generated to increase expression of the encoded polypeptide. In some embodiments, polynucleotide variants are generated to decrease expression of the encoded polypeptide. In some embodiments, polynucleotide variants increase expression of the encoded polypeptide compared to the parent polynucleotide sequence. In some embodiments, polynucleotide variants decrease expression of the encoded polypeptide compared to the parent polynucleotide sequence.

[0181]

[0189] In some embodiments, a polynucleotide comprises a coding sequence for a polypeptide (e.g., an antibody) fused in the same reading frame to a polynucleotide that aids in the expression and secretion of the polypeptide from a host cell (e.g., a leader sequence that functions as a secretory sequence to control transport of the polypeptide). The polypeptide may have a leader sequence that is cleaved by the host cell to form a "mature" form of the polypeptide.

[0182]

[0190] In some embodiments, the polynucleotide comprises a coding sequence for a polypeptide (e.g., an antibody) fused in the same reading frame to a marker or tag sequence. For example, in some embodiments, the marker sequence is a hexa-histidine tag (HIS-tag), which allows for efficient purification of the polypeptide fused to the marker. In some embodiments, when a mammalian host (e.g., COS-7 cells) is used, the marker sequence is a hemagglutinin (HA) tag, derived from the influenza hemagglutinin protein. In some embodiments, the marker sequence is a FLAG™ tag. In some embodiments, the marker can be used in conjunction with other markers or tags.

[0183]

[0191] In some embodiments, the polynucleotide is isolated. In some embodiments, the polynucleotide is substantially pure.

[0192] Also provided are vectors and cells comprising the polynucleotides described herein. In some embodiments, vectors comprising the polynucleotides provided herein are provided. The vectors may be expression vectors. In some embodiments, the vectors provided herein comprise a polynucleotide encoding an anti-MASP-2 antibody or antigen-binding fragment described herein. In some embodiments, the vectors provided herein comprise a polynucleotide encoding a polypeptide that is part of an anti-MASP-2 antibody or antigen-binding fragment described herein.

[0184]

[0193] In some embodiments, recombinant expression vectors are provided that can be used to amplify and express polynucleotides encoding the anti-MASP-2 antibodies or antigen-binding fragments described herein. For example, recombinant expression vectors can be replicable DNA constructs containing synthetic or cDNA-derived DNA fragments encoding polypeptide chains of anti-MASP-2 antibodies operably linked to suitable transcriptional and / or translational regulatory elements derived from mammalian, microbial, viral, or insect genes. In some embodiments, viral vectors are used. DNA regions are "operably linked" when they are functionally related to each other. For example, a promoter is operably linked to a coding sequence if it controls the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned to allow translation. In some embodiments, structural elements intended for use in a particular expression system include a leader sequence enabling extracellular secretion of translated protein by the host cell. In some embodiments, when the recombinant protein is expressed without a leader or transport sequence, the polypeptide may include an N-terminal methionine residue.

[0185]

[0194] A wide variety of expression host / vector combinations can be utilized. Useful expression vectors for eukaryotic hosts include, for example, vectors containing expression control sequences from SV40, bovine papillomavirus, adenovirus, and cytomegalovirus. Useful expression vectors for bacterial hosts include known bacterial plasmids such as E. coli-derived plasmids including pCR1, pBR322, pMB9, and their derivatives, as well as broader host range plasmids such as M13 and other filamentous single-stranded DNA phages.

[0186]

[0195] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein are expressed from one or more vectors. Suitable host cells for expression include prokaryotes, yeast cells, insect cells, or higher eukaryotic cells under the control of a suitable promoter. Suitable cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian hosts, as well as methods for protein production, including antibody production, are well known in the art.

[0187]

[0196] Examples of suitable mammalian host cell lines include, but are not limited to, COS-7 (derived from monkey kidney), L-929 (derived from mouse fibroblasts), C127 (derived from mouse mammary tumor), 3T3 (derived from mouse fibroblasts), CHO (derived from Chinese hamster ovary), HeLa (derived from human cervical carcinoma), BHK (derived from hamster kidney fibroblasts), and HEK-293 (derived from human embryonic kidney) cell lines, and their variants. Mammalian expression vectors may contain nontranscribed elements such as an origin of replication, a suitable promoter and enhancer linked to the gene to be expressed, and other 5'- or 3'-flanking nontranscribed sequences, as well as necessary 5'- or 3'-untranslated sequences such as ribosome binding sites, polyadenylation sites, splice donor and acceptor sites, and transcription termination sequences. Expression of recombinant proteins in insect cell culture systems (e.g., baculovirus) also provides a robust method for producing correctly folded and biologically functional proteins. Baculovirus systems for production of heterologous proteins in insect cells are well known to those skilled in the art.

[0188]

[0197] The present disclosure also provides host cells comprising a polypeptide described herein, a polynucleotide encoding a polypeptide described herein, or a vector comprising such a polynucleotide. In some embodiments, host cells are provided comprising a vector comprising a polynucleotide disclosed herein. In some embodiments, the host cells provided herein comprise a vector comprising a polynucleotide encoding an anti-MASP-2 antibody or antigen-binding fragment described herein. In some embodiments, the host cells provided herein comprise a vector comprising a polynucleotide encoding a polypeptide that is part of the anti-MASP-2 antibody or antigen-binding fragment described herein. In some embodiments, the host cells provided herein comprise a polynucleotide encoding the anti-MASP-2 antibody or antigen-binding fragment described herein. In some embodiments, the cells produce the anti-MASP-2 antibody or antigen-binding fragment described herein. 5.5 Method of production

[0198] The polynucleotides provided herein can be prepared, manipulated, and / or expressed using any of the well-established techniques known and available in the art. Many vectors can be used. Examples of vectors include plasmids, autonomously replicating sequences, and transposable elements. Exemplary transposon systems, such as Sleeping Beauty and PiggyBac, which can be stably integrated into a genome, can be used (e.g., Ivic et al., Cell, 91(4): 501-510(1997); Cadinano et al., (2007) Nucleic Acids Research. 35(12): e87). Further exemplary vectors include, but are not limited to, plasmids, phagemids, cosmids, artificial chromosomes such as yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), or P1-derived artificial chromosomes (PACs), bacteriophages such as lambda phage or M13 phage, and animal viruses. Examples of categories of animal viruses useful as vectors include, but are not limited to, retroviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpes viruses (e.g., herpes simplex virus), poxviruses, baculoviruses, papillomaviruses, and papovaviruses (e.g., SV40). Exemplary expression vectors include the pClneo vector (Promega) for mammalian expression, and pLenti4 / V5-DEST™, pLenti6 / V5-DEST™, and pLenti6.2 / V5-GW / lacZ (Invitrogen) for lentivirus-mediated gene transfer and expression in mammals.

[0189]

[0199] In some embodiments, the vector is an episomal vector or a vector that is maintained extrachromosomally. As used herein, the term "episomal" refers to a vector that can replicate without integrating into the host's chromosomal DNA and without being gradually lost from dividing host cells, and also means that the vector replicates extrachromosomally or episomally. The vector is modified to carry a sequence encoding a DNA origin of replication or "ori" from a lymphotropic herpesvirus or gammaherpesvirus, adenovirus, SV40, bovine papillomavirus, or yeast, specifically, the lymphotropic herpesvirus or gammaherpesvirus replication origin corresponding to the oriP of EBV. In some embodiments, the lymphotropic herpesvirus can be Epstein-Barr virus (EBV), Kaposi's sarcoma herpesvirus (KSHV), herpesvirus saimiri (HS), or Marek's disease virus (MDV). Epstein-Barr virus (EBV) and Kaposi's sarcoma herpesvirus (KSHV) are also examples of gammaherpesviruses. Typically, host cells contain viral replication transactivator proteins that activate replication.

[0190]

[0200] "Expression control sequences," "control elements," or "regulatory sequences" present in an expression vector are the untranslated regions of the vector—origins of replication, selection cassettes, promoters, enhancers, translation initiation signals (Shine-Dalgarno or Kozak sequences), introns, polyadenylation sequences, and 5' and 3' untranslated regions—that interact with host cellular proteins to carry out transcription and translation. Such elements can vary in strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including ubiquitous and inducible promoters, can be used.

[0191]

[0201] Exemplary ubiquitous expression control sequences that can be used in the present disclosure include the cytomegalovirus (CMV) immediate early promoter, the viral simian virus 40 (SV40) promoter (e.g., early or late), the Moloney murine leukemia virus (MoMLV) LTR promoter, the Rous sarcoma virus (RSV) LTR, the herpes simplex virus (HSV) (thymidine kinase) promoter, the H5, P7.5, and P11 promoters from vaccinia virus, the elongation factor 1-alpha (EF1a) promoter, the early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), glyceraldehyde-3-methyltransferase (GMT) promoter, and the glyceraldehyde-3-methyltransferase (GMT) promoter. Examples of suitable promoters include, but are not limited to, 3-phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), heat shock 70 kDa protein 5 (HSPA5), heat shock protein 90 kDa β, member 1 (HSP90B1), heat shock protein 70 kDa (HSP70), β-kinesin (β-KIN), human ROSA 26 locus (Irion et al., Nature Biotechnology 25, 1477-1482 (2007)), ubiquitin C promoter (UBC), phosphoglycerate kinase-1 (PGK) promoter, cytomegalovirus enhancer / chicken β-actin (CAG) promoter, and β-actin promoter.

[0192]

[0202] Examples of inducible promoters / systems include, but are not limited to, steroid-inducible promoters, such as promoters of genes encoding glucocorticoid or estrogen receptors (induced by treatment with the corresponding hormones), metallothionein promoters (induced by treatment with various heavy metals), MX-1 promoters (induced by interferon), the "GeneSwitch" mifepristone-regulatable system (Sirinet et al., 2003, Gene, 323:67), cumate-inducible gene switches (WO 2002 / 088346), and tetracycline-dependent regulatory systems. The anti-MASP-2 antibodies or antigen-binding fragments described herein can be produced by any method known in the art, including chemical synthesis and recombinant expression techniques. The practice of the present invention utilizes, unless otherwise indicated, conventional techniques in molecular biology, microbiology, genetic analysis, recombinant DNA, organic chemistry, biochemistry, PCR, oligonucleotide synthesis and modification, nucleic acid hybridization, and related fields within the skill of the art. These techniques are described in the references cited herein and are fully explained in the literature, e.g., Maniatis et al., (1982), Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Sambrook et al., (1989), Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press; Sambrook et al., (2001), Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Ausubel et al.Current Protocols in Molecular Biology, John Wiley & Sons (1987 and annually updated editions); Current Protocols in Immunology, John Wiley & Sons (1987 and annually updated editions), Gait (ed.) (1984), Oligonucleotide Synthesis: A Practical Approach, IRL Press; Eckstein (ed.) (1991), Oligonucleotides and Analogues: A Practical Approach, IRL Press; Birren et al., (eds.) (1999), Genome Analysis: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Borrebaeck (ed.) (1995), Antibody Engineering, 2nd ed., Oxford University Press; Lo (ed.) (2006), Antibody Engineering: Methods and Protocols See Methods in Molecular Biology, Vol. 248, Humana Press, 1999, pp. 248-252, each of which is incorporated herein by reference in its entirety.

[0193]

[0203] The polypeptides described herein (e.g., anti-MASP-2 antibodies or antigen-binding fragments) can be produced and isolated using methods known in the art. Peptides can also be synthesized in whole or in part using chemical methods (see, e.g., Caruthers (1980). Nucleic Acids Res. Symp. Ser. 215; Horn (1980); and Banga, A.K., Therapeutic Peptides and Proteins, Formulation, Processing and Delivery Systems (1995) Technomic Publishing Co., Lancaster, Pa.). Peptide synthesis can be carried out using a variety of solid-phase methods (see, e.g., Roberge, Science 269:202 (1995); Merrifield, Methods. Enzymol. 289:3 (1997)), and automated synthesis can be achieved using, for example, an ABI 431A peptide synthesizer (Perkin Elmer) according to the manufacturer's instructions. Peptides can also be synthesized using combinatorial methods. Synthetic residues and polypeptides can be synthesized using a variety of procedures and methods known in the art (see, e.g., Organic Syntheses Collective Volumes, Gilman et al., (eds.) John Wiley & Sons, Inc., NY). Modified peptides can be produced by chemical modification methods (see, e.g., Belousov, Nucleic Acids Res. 25:3440 (1997); Frenkel, Free Radic. Biol. Med. 19:373 (1995); and Blommers, Biochemistry 33:7886 (1994)). Peptide sequence variations, derivatives, substitutions, and modifications can also be made using methods such as oligonucleotide-mediated (site-directed) mutagenesis, alanine scanning, and PCR-based mutagenesis.Site-directed mutagenesis (Carter et al., Nucl. Acids Res., 13:4331 (1986); Zoller et al., Nucl. Acids Res. 10:6487 (1987)), cassette mutagenesis (Wells et al., Gene 34:315 (1985)), restriction-selection mutagenesis (Wells et al., Philos. Trans. R. Soc. London SerA 317:415 (1986)), and other techniques can be performed on the cloned DNA to produce inventive peptide sequences, variants, fusions, and chimeras, as well as mutations, derivatives, substitutions, and modifications thereof.

[0194]

[0204] The polypeptides described herein can be prepared using a variety of techniques known in the art, including the use of hybridoma and recombinant technologies, or a combination thereof. In some embodiments, a recombinant expression vector is used to express a polynucleotide encoding a polypeptide described herein. For example, a recombinant expression vector can be a replicable DNA construct comprising a synthetic or cDNA-derived DNA fragment encoding a polypeptide operably linked to suitable transcriptional and / or translational regulatory elements derived from mammalian, microbial, viral, or insect genes. In some embodiments, the coding sequence of a polypeptide disclosed herein can be ligated to such an expression vector for its expression in a mammal. In some embodiments, viral vectors are used. DNA regions are "operably linked" when they are functionally related to each other. For example, a promoter is operably linked to a coding sequence if it controls the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to permit translation. In some embodiments, structural elements intended for use in yeast expression systems include a leader sequence enabling extracellular secretion of translated protein by the host cell. In some embodiments, when the recombinant protein is expressed without a leader or transport sequence, the polypeptide may include an N-terminal methionine residue.

[0195]

[0205] A wide variety of expression host / vector combinations are available. Suitable host cells for expression include prokaryotes, yeast cells, insect cells, or higher eukaryotic cells under the control of an appropriate promoter. Suitable cloning and expression vectors for use with bacterial, fungal, yeast, and mammalian cell hosts, as well as methods for protein production, including antibody production, are well known in the art. Useful expression vectors for bacterial hosts include known bacterial plasmids such as E. coli-derived plasmids, including pCR1, pBR322, pMB9, and their derivatives, as well as broader host range plasmids such as M13 and other filamentous single-stranded DNA phages.

[0196]

[0206] Useful expression vectors for eukaryotic hosts include, for example, vectors containing expression control sequences derived from SV40, bovine papilloma virus, adenovirus, and cytomegalovirus. Examples of suitable mammalian host cell lines include, but are not limited to, COS-7 (derived from monkey kidney), L-929 (derived from mouse fibroblast), C127 (derived from mouse mammary tumor), 3T3 (derived from mouse fibroblast), CHO (derived from Chinese hamster ovary), HeLa (derived from human cervical carcinoma), BHK (derived from hamster kidney fibroblast), HEK-293 (derived from human embryonic kidney) cell lines, and variants thereof. Mammalian expression vectors may include nontranscribed elements such as an origin of replication, a suitable promoter and enhancer linked to the gene to be expressed, and other 5'- or 3'-flanking nontranscribed sequences, as well as necessary ribosome binding sites, polyadenylation sites, splice donor and acceptor sites, and transcription termination sequences. Expression of recombinant proteins in insect cell culture systems (e.g., baculovirus) also provides a robust method for producing correctly folded and biologically functional proteins. Baculovirus systems for the production of heterologous proteins in insect cells are well known to those skilled in the art.

[0197]

[0207] Provided herein are anti-MASP-2 antibodies and antigen-binding fragments thereof, including but not limited to monoclonal antibodies, polyclonal antibodies, synthetic antibodies, human antibodies, humanized antibodies, and antigen-binding fragments thereof.

[0198]

[0208] Methods for producing antibodies are well known in the art. See, for example, Harlow et al., Antibodies: A Laboratory Manual (Cold Spring Harbor Laboratory Press, 2nd ed., 1988); Hammerling et al., Monoclonal Antibodies and T-Cell Hybridomas, 563-681 (Elsevier, NY, 1981). Each of these references is incorporated herein by reference in its entirety. For in vivo use of antibodies in humans, it may be preferable to use human antibodies. For therapeutic treatment of human subjects, fully human antibodies are particularly desirable. Human antibodies can be produced by a variety of methods known in the art, including phage display methods using antibody libraries derived from human immunoglobulin sequences, along with modifications of these techniques. See also U.S. Patent Nos. 4,444,887 and 4,716,111; and PCT Publication Nos. WO 98 / 46645, WO 98 / 50433, WO 98 / 24893, WO 98 / 16654, WO 96 / 34096, WO 96 / 33735, and WO 91 / 10741, each of which is incorporated herein by reference in its entirety. A human antibody can also be an antibody in which the heavy and light chains are encoded by nucleotide sequences derived from one or more sources of human DNA.

[0199]

[0209] Human antibodies can also be produced using transgenic mice that are incapable of expressing functional endogenous immunoglobulins but can express human immunoglobulin genes. For example, human heavy and light chain immunoglobulin gene complexes can be introduced randomly or by homologous recombination into mouse embryonic stem cells. Alternatively, human variable, constant, and diversity regions can be introduced into mouse embryonic stem cells in addition to human heavy and light chain genes. The mouse heavy and light chain immunoglobulin genes can be rendered nonfunctional separately or simultaneously with the introduction of human immunoglobulin loci by homologous recombination. For example, it has been described that homozygous deletion of the antibody heavy chain joining region (JH) gene in chimeric or germline mutant mice completely abolishes endogenous antibody production. These modified embryonic stem cells are expanded and microinjected into blastocysts to generate chimeric mice. The chimeric mice are then bred to produce homozygous offspring that express human antibodies. The transgenic mice are immunized in the usual manner with a selected antigen, such as all or a portion of a polypeptide of the invention. For example, anti-MASP-2 antibodies against the human MASP-2 antigen can be obtained from immunized transgenic mice using conventional hybridoma technology. The human immunoglobulin transgenes carried by the transgenic mice rearrange during B cell differentiation and subsequently undergo class switching and somatic mutation. Thus, using such technology, it is possible to produce therapeutically useful IgG, IgA, IgM, and IgE antibodies, including, but not limited to, IgG1 (γ1) and IgG3. For an overview of this technology for producing human antibodies, see Lonberg and Huszar (Int. Rev. Immunol, 13:65-93 (1995)).For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and protocols for producing such antibodies, see, e.g., PCT Publication Nos. WO 98 / 24893, WO 96 / 34096, and WO 96 / 33735; and U.S. Patent Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,806; 5,814,318; and 5,939,598. Each of these patents is incorporated herein by reference in its entirety. Additionally, companies such as Abgenix, Inc. (Freemont, Calif.) and Genpharm (San Jose, Calif.) can be contracted to provide human antibodies directed against a selected antigen using technology similar to that described above. For specific discussion of the transfer of human germ-line immunoglobulin gene arrays into germ-line mutant mice, which would result in the production of human antibodies upon antigen challenge, see, e.g., Jakobovits et al., Proc. Natl. Acad. Sci. USA, 90:2551 (1993); Jakobovits et al., Nature, 362:255-258 (1993); Bruggermann et al., Year in Immunol, 7:33 (1993); and Duchosal et al., Nature, 355:258 (1992).

[0200]

[0210] Human antibodies can also be derived from phage display libraries (Hoogenboom et al., J. Mol. Biol, 227:381 (1991); Marks et al., J. Mol. Biol, 222:581-597 (1991); Vaughan et al., Nature Biotech., 14:309 (1996)). Using phage display technology (McCafferty et al., Nature, 348:552-553 (1990)), human antibodies and antibody fragments can be produced in vitro from immunoglobulin variable (V) domain gene repertoires of unimmunized donors. According to this technique, antibody V domain genes are cloned in frame into either a major or minor coat protein gene of a filamentous bacteriophage, such as M13 or fd, and displayed as functional antibody fragments on the surface of the phage particle. Because the filamentous particle contains a single-stranded DNA copy of the phage genome, selection based on the functional properties of the antibody also results in selection of the gene encoding the antibody exhibiting those properties. Thus, the phage mimics some of the properties of B cells. Phage display can be performed in a variety of formats. For reviews, see, e.g., Johnson and Chiswell, Current Opinion in Structural Biology 3:564-571 (1993). Several sources of V gene segments can be used for phage display. Clackson et al., Nature, 352:624-628 (1991) isolated a diverse array of anti-oxazolone antibodies from a small random combinatorial library of V genes derived from the spleens of immunized mice.Repertoires of V genes from unimmunized human donors can be constructed, and antibodies to a diverse array of antigens (including self-antigens) can be isolated essentially according to the techniques described in Marks et al., J. Mol. Biol, 222:581-597 (1991), or Griffith et al., EMBO J., 12:725-734 (1993). See also U.S. Patent Nos. 5,565,332 and 5,573,905, each of which is incorporated herein by reference in its entirety.

[0201]

[0211] Human antibodies can also be produced by in vitro activated B cells (see U.S. Patent Nos. 5,567,610 and 5,229,275, each of which is incorporated herein by reference in its entirety). Human antibodies can also be produced in vitro using hybridoma techniques, such as, but not limited to, those described by Roder et al., (Methods Enzymol., 121:140-167 (1986)).

[0202]

[0212] Alternatively, in some embodiments, non-human antibodies are humanized, in which specific sequences or regions of the antibody are modified to increase their similarity to antibodies naturally produced in humans, hi some embodiments, the antigen-binding domain portion is humanized.

[0203]

[0213] Humanized antibodies can be produced using various techniques known in the art, including, but not limited to, CDR grafting (see, e.g., European Patent No. EP 239,400; International Publication No. WO 91 / 09967; and U.S. Pat. Nos. 5,225,539, 5,530,101, and 5,585,089, each of which is incorporated herein by reference in its entirety), veneering, or resurfacing (see, e.g., European Patent Nos. EP 592,106 and EP 519,596; Padlan, 1991, Molecular Immunology 28(4 / 5):489-498; Studnicka et al., 1994, Protein Engineering 7(6):805-814; and Roguska et al., 1994, Proc. See Natl Acad Sci USA 91:969-973, each of which is incorporated herein by reference in its entirety), chain shuffling (see, e.g., U.S. Pat. No. 5,565,332).No. 6,407,213, U.S. Pat. No. 5,766,886, International Publication No. WO 9317105, Tan et al., J. Immunol., 169: 1119-25 (2002), Caldas et al., Protein Eng., 13(5):353-60 (2000), Morea et al., Methods, 20(3):267-79 (2000), Baca et al., J. Biol. Chem., 272(16):10678-84 (1997), Roguska et al., Protein Eng., 9(10):895-904 (1996), Couto et al., Cancer Res., 55 (23 Supp):5973s-5977s (1995), Couto et al., Cancer Res., 55(8): 1717-22 (1995), Sandhu JS, Gene, 150(2):409-10 (1994), and Pedersen et al., J. Mol. Biol., 235(3):959-73 (1994), each of which is incorporated herein by reference in its entirety. In many cases, framework residues in the framework regions can be substituted with corresponding residues from the CDR donor antibody to alter, preferably improve, antigen binding. These framework substitutions are identified by methods well known in the art, such as by modeling the interactions of CDRs and framework residues to identify framework residues important for antigen binding, and by sequence comparison to identify unusual framework residues at specific positions (see, e.g., Queen et al., U.S. Pat. No. 5,585,089; and Riechmann et al., 1988, Nature, 332:323; each of these patent documents is incorporated herein by reference in its entirety).

[0204]

[0214] A humanized antibody has one or more amino acid residues introduced into it from a non-human source. These non-human amino acid residues, often referred to as "import" residues, are typically taken from an "import" variable domain. A humanized antibody thus comprises one or more CDRs from a non-human immunoglobulin molecule and a framework region of human origin. Antibody humanization is well known in the art and essentially involves substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody, i.e., CDR grafting (EP 239,400; PCT Publication No. WO 2014 / 023106). 91 / 09967; and U.S. Patent Nos. 4,816,567; 6,331,415; 5,225,539; 5,530,101; 5,585,089; and 6,548,640, the contents of which are incorporated herein by reference in their entireties. In such humanized chimeric antibodies, substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some framework (FR) residues are substituted by residues from analogous sites in rodent antibodies. Antibody humanization can also be achieved by veneering or resurfacing (EP 592,106; EP 519,596; Padlan, 1991, Molecular Immunology, 28(4 / 5):489-498; Studnicka et al., Protein Engineering, 7(6):805-814 (1994); and Roguska et al., PNAS, 91:969-973 (1994)) or chain shuffling (U.S. Pat. No. 5,565,332), the contents of which are incorporated herein by reference in their entireties.

[0205]

[0215] The human variable domains, both light and heavy, used to create humanized antibodies are selected to reduce antigenicity. According to the so-called "best-fit" method, the sequence of the variable domain of a rodent antibody is screened against the entire library of known human variable domain sequences. The human sequence that is closest to the rodent sequence is then accepted as the human framework (FR) of the humanized antibody (Sims et al., J. Immunol., 151:2296 (1993); Chothia et al., J. Mol. Biol., 196:901 (1987), the contents of which are incorporated herein by reference in their entirety). Another method uses a specific framework derived from the consensus sequence of all human antibodies of a particular light or heavy chain subgroup. The same framework can be used for several different humanized antibodies (Carter et al., Proc. Natl. Acad. Sci. USA, 89:4285 (1992); Presta et al., J. Immunol., 151:2623 (1993), the contents of which are incorporated herein by reference in their entireties).

[0206]

[0216] Antibodies can be humanized while retaining high affinity for the target antigen and other favorable biological properties. For example, humanized antibodies can be prepared by analyzing the parental sequences and various conceptual humanized products using three-dimensional models of the parental and humanized sequences. Three-dimensional immunoglobulin models are publicly available and familiar to those skilled in the art. Computer programs are available that illustrate and display probable three-dimensional conformations of selected candidate immunoglobulin sequences. Inspection of these displays permits analysis of the role of the residues in the function of the candidate immunoglobulin sequence, i.e., analysis of residues that influence the ability of the candidate immunoglobulin to bind to the target antigen. In this way, FR residues from the recipient and import sequences can be selected and combined to achieve desired antibody characteristics, such as improved affinity for the target antigen. In general, CDR residues are directly and most substantially involved in influencing antigen binding.

[0207]

[0217] Humanized antibodies retain the same antigen specificity as the original antibody, such as the ability to bind to the human MASP-2 antigen. However, certain humanization methods can be used to increase the binding affinity and / or specificity of an antibody for a particular antigen using methods of "directed evolution," such as those described by Wu et al., J. Mol. Biol., 294:151 (1999), the entire contents of which are incorporated herein by reference. 5.6 Pharmaceutical Compositions

[0218] Also provided herein are pharmaceutical compositions comprising the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein. In some embodiments, the pharmaceutical compositions comprise a therapeutically effective amount of the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions are useful in inhibiting MASP-2-dependent complement activation. In some embodiments, the pharmaceutical compositions are useful in treating diseases or disorders associated with MASP-2-dependent complement activation.

[0208]

[0219] In some embodiments, the pharmaceutical compositions provided herein comprise an anti-MASP-2 antibody or antigen-binding fragment provided herein. The anti-MASP-2 antibody or antigen-binding fragment may be present at various concentrations. In some embodiments, the pharmaceutical compositions provided herein comprise a soluble anti-MASP-2 antibody or antigen-binding fragment provided herein at 1 to 1000 mg / mL. In some embodiments, the pharmaceutical compositions comprise a soluble anti-MASP-2 antibody or antigen-binding fragment provided herein at 10 to 500 mg / mL, 10 to 400 mg / mL, 10 to 300 mg / mL, 10 to 200 mg / mL, 10 to 100 mg / mL, 20 to 100 mg / mL, or 50 to 100 mg / mL. In some embodiments, the pharmaceutical compositions provided herein contain an anti-MASP-2 antibody or antigen-binding fragment provided herein at about 10 mg / mL, about 20 mg / mL, about 30 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 150 mg / mL, about 180 mg / mL, about 200 mg / mL, about 300 mg / mL, about 500 mg / mL, about 800 mg / mL, or about 1000 mg / mL. The dosage can be easily adjusted by one skilled in the art; for example, a decrease in purity may require an increased dosage.

[0209]

[0220] Also provided herein are kits for preparing pharmaceutical compositions having the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein. In some embodiments, the kits comprise the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein and a pharmaceutically acceptable carrier in one or more containers. In another embodiment, the kits may comprise the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein for administration to a subject. In certain embodiments, the kits include instructions for preparing and / or administering the anti-MASP-2 antibodies or antigen-binding fragments.

[0210]

[0221] In some embodiments, a pharmaceutical composition is provided comprising the anti-MASP-2 antibody or antigen-binding fragment or cell provided herein, which composition is suitable for topical administration.

[0211]

[0222] Pharmaceutically acceptable carriers that may be used in the compositions provided herein include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. In some embodiments, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal, or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active ingredient (i.e., anti-MASP-2 antibody or antigen-binding fragment) may be coated with a material to protect the active ingredient from the action of acids and other natural conditions that may inactivate the active ingredient.

[0212]

[0223] Also provided herein are pharmaceutical compositions or formulations that improve the stability of anti-MASP-2 antibodies or antigen-binding fragments to enable long-term storage of the antibodies or antigen-binding fragments. In some embodiments, the pharmaceutical compositions or formulations disclosed herein comprise (a) an anti-MASP-2 antibody or antigen-binding fragment disclosed herein, (b) a buffer, (c) a stabilizer, (d) a salt, (e) a bulking agent, and / or (f) a surfactant. In some embodiments, the pharmaceutical composition or formulation is stable for at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 1 year, at least 2 years, at least 3 years, at least 5 years, or longer. In some embodiments, the pharmaceutical composition or formulation is stable when stored at 4°C, 25°C, or 40°C.

[0213]

[0224] Buffers useful in the pharmaceutical compositions or formulations disclosed herein can be weak acids or bases that maintain the acidity (pH) of a solution near a selected value after the addition of another acid or base. A suitable buffering agent can maximize the stability of a pharmaceutical formulation by maintaining pH control of the formulation. A suitable buffering agent can also ensure physiological compatibility or optimize solubility. Rheology, viscosity, and other properties can also depend on the pH of the formulation. Common buffering agents include, but are not limited to, histidine, citrate, succinate, acetate, and phosphate. In some embodiments, the buffering agent includes histidine (e.g., L-histidine) along with an isotonicity agent, potentially with pH adjustment by an acid or base known in the art. In certain embodiments, the buffering agent is L-histidine. In certain embodiments, the pH of the formulation is maintained at about 2 to about 10, or about 4 to about 8.

[0214]

[0225] Stabilizers are added to pharmaceutical products to stabilize them. Such agents can stabilize proteins in various ways. Common stabilizers include, but are not limited to, amino acids such as glycine, alanine, lysine, arginine, or threonine; carbohydrates such as glucose, sucrose, trehalose, raftose, or maltose; polyols such as glycerol, mannitol, sorbitol, cyclodextrins or destranes of any type and molecular weight, or PEG. In some embodiments, the stabilizer is selected to maximize the stability of the FIX polypeptide in the lyophilized preparation. In certain embodiments, the stabilizer is sucrose and / or arginine.

[0215]

[0226] Bulking agents may be added to pharmaceutical compositions or formulations to add volume and mass to the product, thereby facilitating its accurate measurement and handling. Common bulking agents include, but are not limited to, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate, or magnesium stearate.

[0216]

[0227] Surfactants are amphiphilic substances having a hydrophilic group and a hydrophobic group. Surfactants can be anionic, cationic, zwitterionic, or nonionic. Examples of nonionic surfactants include, but are not limited to, alkyl ethoxylates, nonylphenol ethoxylates, amine ethoxylates, polyethylene oxide, polypropylene oxide, fatty alcohols such as cetyl alcohol or oleyl alcohol, cocamide MEA, cocamide DEA, polysorbates, or dodecyldimethylamine oxide. In some embodiments, the surfactant is polysorbate 20 or polysorbate 80.

[0217]

[0228] The pharmaceutical compositions disclosed herein may further comprise one or more of a buffer system, a preservative, a tonicity agent, a chelating agent, a stabilizer, and / or a surfactant, as well as various combinations thereof. The use of preservatives, isotonicity agents, chelating agents, stabilizers, and surfactants in pharmaceutical compositions is well known to those skilled in the art. See Remington: The Science and Practice of Pharmacy, 19th Edition, 1995.

[0218]

[0229] In some embodiments, the pharmaceutical composition is an aqueous formulation. Such formulations are typically solutions or suspensions, but may also include colloids, dispersions, emulsions, and multiphase materials. The term "aqueous formulation" is defined as a formulation that contains at least 50% w / w water. Similarly, the term "aqueous solution" is defined as a solution that contains at least 50% w / w water, and the term "aqueous suspension" is defined as a suspension that contains at least 50% w / w water.

[0219]

[0230] In some embodiments, the pharmaceutical compositions disclosed herein are lyophilized, and the physician or the patient adds solvents and / or diluents thereto prior to use.

[0231] The pharmaceutical compositions disclosed herein may also contain a pharmaceutically acceptable antioxidant. Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc., (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc., and (3) metal chelators, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0220]

[0232] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions or formulations described herein include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0221]

[0233] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the presence of microorganisms can be ensured by both the above-mentioned sterilization procedures and the inclusion of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include isotonic agents, such as sugars and sodium chloride, in the compositions. Furthermore, the inclusion of agents that delay absorption, such as aluminum monostearate and gelatin, can prolong the absorption of injectable pharmaceutical forms.

[0222]

[0234] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions.The use of such media and agents for pharmaceutically active substances is well known in the art.Except where a conventional media or agent is incompatible with the active compound, its use in the pharmaceutical compositions described herein is contemplated.The pharmaceutical compositions or preparations may contain or be devoid of preservatives.A supplementary active compound may be incorporated into this composition.

[0223]

[0235] Pharmaceutical compositions or formulations must typically be sterile and stable under the conditions of manufacture and storage. The composition may be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, and the like), and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. In many cases, these compositions may contain isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. The absorption of injectable compositions can be prolonged by including in the composition an agent that delays absorption, such as monostearate salts and gelatin.

[0224]

[0236] Sterile injection solution can be prepared by incorporating the active compound in the required amount in a suitable solvent with one or a combination of the above-mentioned components, and then optionally sterilizing by microfiltration.Usually, dispersion is prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and other components required from the components listed in this specification.For the preparation of sterile powder for sterile injection solution, some methods are vacuum drying and freeze-drying (lyophilization), which obtains the powder of active ingredient and any other desired components from the solution that has been previously sterilized and filtered.

[0225]

[0237] The amount of active ingredient that can be combined with a carrier material in a pharmaceutical composition or formulation disclosed herein can vary. In some embodiments, the amount of active ingredient that can be combined with a carrier material is an amount that produces a therapeutic effect. Typically, out of 100%, this amount ranges from about 0.01% to about 99% of the active ingredient combined with a pharmaceutically acceptable carrier, from about 0.1% to about 70% of the active ingredient, or from about 1% to about 30%.

[0226]

[0238] The pharmaceutical compositions disclosed herein may be prepared using carriers that protect the active ingredient from rapid release, such as controlled-release formulations, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid may be used. Many methods for preparing such formulations are patented or generally known to those skilled in the art. See, for example, "Sustained and Controlled Release Drug Delivery Systems," edited by JR Robinson, Marcel Dekker, New York, 1978.

[0227]

[0239] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein can be formulated to ensure proper distribution in vivo. For example, the blood-brain barrier (BBB) excludes many highly hydrophilic compounds. To ensure that the active ingredients described herein cross the BBB, they can be formulated, for example, in liposomes. For methods of manufacturing liposomes, see, for example, U.S. Patent Nos. 4,522,811, 5,374,548, and 5,399,331. Liposomes can contain one or more moieties that selectively transport to specific cells or organs, thereby enhancing targeted drug delivery (see, e.g., V. V. Ranade (1989) J. Clin. Pharmacol. 29:685). Exemplary targeting moieties include folate or biotin (see, e.g., U.S. Patent No. 5,416,016 to Low et al.), mannosides (Umezawa et al., (1988) Biochem. Biophys. Res. Commun. 153: 1038); antibodies (P.G. Bloeman et al., (1995) FEBS Lett. 357: 140; M. Owais et al., (1995) Antimicrob. Agents Chemother. 39: 180); surfactant protein A receptor (Briscoe et al., (1995) Am. J. Physiol. 1233: 134); p120 (Schreier et al., (1994) J. Biol. Chem. 269:9090); K. Keinanen; M.L. Laukkanen (1994) FEBS Lett. 346:123; see also JJ Killion; IJ Fidler (1994) Immunomethods 4:273.

[0228]

[0240] The anti-MASP-2 antibodies or antigen-binding fragments described herein can be tested for binding to human MASP-2, for example, by standard ELISA. Briefly, microtiter plates are coated with purified MASP-2 and then blocked with bovine serum albumin. Dilutions of antibody (e.g., dilutions of plasma from MASP-2-immunized mice) are added to each well and incubated. The plate is washed and incubated with a secondary reagent conjugated to horseradish peroxidase (HRP) (e.g., in the case of human antibodies, a goat anti-human IgG Fc-specific polyclonal reagent). After washing, the plate can be developed and analyzed by spectrophotometry. Serum from immunized mice can then be further screened by flow cytometry for binding to cell lines expressing human MASP-2, but not to control cell lines that do not express MASP-2. Briefly, binding of anti-MASP-2 antibodies can be assessed by incubating MASP-2-expressing CHO cells with the anti-MASP-2 antibody. Cells can be washed and binding can be detected with anti-human IgG Ab. Flow cytometric analysis can be performed using a FACScan flow cytometer (Becton Dickinson, San Jose, CA). Mice developing the highest titers can be used for fusions.

[0229]

[0241] The above-described ELISA assay can be used to screen for antibodies, and thus hybridomas that produce antibodies that show positive reactivity with the MASP-2 immunogen. Hybridomas that produce antibodies that bind to MASP-2 with high affinity can then be subcloned and further characterized. One clone from each hybridoma that retains the reactivity of the parent cells (by ELISA) can then be selected for cell banking and antibody purification.

[0230]

[0242] To purify anti-MASP-2 antibodies, selected hybridomas can be expanded for monoclonal antibody purification. The supernatant can be filtered and concentrated before affinity chromatography. To ensure purity, the eluted IgG can be examined by gel electrophoresis and high-performance liquid chromatography. The buffer solution can be exchanged, and the concentration can be determined. The monoclonal antibody can be aliquoted and stored.

[0231]

[0243] To determine whether the selected anti-MASP-2 monoclonal antibodies bind to unique epitopes, each antibody can be biotinylated using commercially available reagents (Pierce, Rockford, IL). Binding of biotinylated MAbs can be detected with a streptavidin-labeled probe. A competition test using unlabeled and biotinylated monoclonal antibodies can be performed using the MASP-2-coated ELISA plate described above.

[0232]

[0244] To determine the isotype of purified antibodies, an isotype ELISA can be performed using reagents specific for antibodies of a particular isotype. For example, to determine the isotype of a human monoclonal antibody, the wells of a microtiter plate can be coated with 1 pg / mL anti-human immunoglobulin overnight at 4°C. After blocking with 1% BSA, the plate is reacted with 1 pg / mL or less of the test monoclonal antibody or purified isotype control for 1-2 hours at ambient temperature. The wells can then be reacted with either a human IgG1- or human IgM-specific alkaline phosphatase-conjugated probe. The plate is developed and analyzed as described above.

[0233]

[0245] To test the binding of monoclonal antibodies to live cells expressing MASP-2, flow cytometry can be used, as described in the Examples. Briefly, cell lines expressing membrane-bound MASP-2 (grown under standard growth conditions) are mixed with various concentrations of monoclonal antibodies in PBS containing 0.1% BSA for 1 hour at 4°C. After washing, the cells are reacted with fluorescein-labeled anti-IgG antibodies under the same conditions as for primary antibody staining. Samples can be gated for single cells using light and side scatter characteristics and analyzed using a FACScan instrument to determine the binding of the labeled antibody. In addition to or instead of flow cytometry assays, another assay using fluorescence microscopy can be used. Cells can be stained exactly as described above and examined by fluorescence microscopy. This method allows visualization of individual cells, but may reduce sensitivity depending on the density of the antigen.

[0234]

[0246] Anti-MASP-2 antibodies or antigen-binding fragments can be further tested for reactivity with the MASP-2 antigen by Western blotting. Briefly, cell extracts from cells expressing MASP-2 can be prepared and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis. After electrophoresis, the separated antigens are transferred to a nitrocellulose membrane, blocked with 20% mouse serum, and probed with the monoclonal antibody to be tested. IgG binding is detected using anti-IgG alkaline phosphatase and developed using BCIP / NBT substrate tablets (Sigma Chem. Co., St. Louis, MO).

[0235]

[0247] Methods for analyzing the binding affinity, cross-reactivity, and binding kinetics of various anti-MASP-2 antibodies include standard assays known in the art, such as biolayer interferometry (BLI) using a Gator system (Probe Life) or an Octet-96 system (Sartorius AG), or BIACORE™ surface plasmon resonance (SPR) analysis using a BIACORE™ 2000 SPR instrument (Biacore AB, Uppsala, Sweden). 5.7 Methods and Uses

[0248] The antibodies or antigen-binding fragments, compositions, and methods described herein have numerous in vitro and in vivo utilities, including, for example, reducing inflammation by inhibiting (or antagonizing) MASP-2-dependent complement activation. The antibodies or antigen-binding fragments provided herein can also be used to detect MASP-2. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments described herein are humanized antibodies or antigen-binding fragments. For example, the anti-MASP-2 antibodies or antigen-binding fragments described herein can be administered to cultured cells in vitro or ex vivo, or can be administered to human subjects in vivo, for example, to selectively inhibit lectin pathway complement activation in various diseases. Thus, provided herein is a method for modifying complement activation in a subject, comprising administering to the subject an anti-MASP-2 antibody or antigen-binding portion thereof described herein, such that complement activation in the subject is modified.

[0236]

[0249] Also included is a method for detecting the presence or amount of human MASP-2 antigen in a sample, comprising contacting the sample and a control sample with a monoclonal antibody, e.g., a humanized monoclonal antibody or its antigen-binding portion, that specifically binds to human MASP-2 under conditions that allow the formation of a complex between the antibody or antigen-binding fragment and human MASP-2. The formation of the complex is then detected, whereby differential complex formation between the sample and the control sample indicates the presence of human MASP-2 antigen in the sample. Furthermore, human MASP-2 can be purified by immunoaffinity purification using the anti-MASP-2 antibodies or antigen-binding fragments described herein.

[0237]

[0250] The present disclosure also provides methods of using the anti-MASP-2 antibodies or antigen-binding fragments disclosed herein, polynucleotides encoding such anti-MASP-2 antibodies or antigen-binding fragments, vectors containing such polynucleotides, or pharmaceutical compositions comprising such antibodies or antigen-binding fragments in the inhibition of MASP-2-dependent complement activation or in the treatment of diseases or disorders associated with MASP-2-dependent complement activation.

[0238]

[0251] In some embodiments, a method is provided for treating a disease or disorder associated with MASP-2-dependent complement activation in a subject in need of treatment, the method comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment disclosed herein. In some embodiments, the use of an anti-MASP-2 antibody or antigen-binding fragment disclosed herein in the treatment of a disease or disorder associated with MASP-2-dependent complement activation is provided. In some embodiments, the use of an anti-MASP-2 antibody or antigen-binding fragment provided herein for the preparation of a medicament for the treatment of a disease or disorder associated with MASP-2-dependent complement activation is provided.

[0239]

[0252] In some embodiments, a method is provided for treating a disease or disorder associated with MASP-2-dependent complement activation in a subject in need of such treatment, the method comprising administering a therapeutically effective amount of a pharmaceutical composition disclosed herein to the subject. In some embodiments, the use of a pharmaceutical composition disclosed herein in the treatment of a disease or disorder associated with MASP-2-dependent complement activation is provided. In some embodiments, the use of a pharmaceutical composition provided herein for the preparation of a medicament for the treatment of a disease or disorder associated with MASP-2-dependent complement activation is provided.

[0240]

[0253] As is known in the art, unregulated, undesired, or excessive activation of the complement system and deposition of immune complexes causes inflammation and damage in various tissues, which are involved in the pathogenesis of various diseases. In some embodiments, the disease or disorder associated with MASP-2-dependent complement activation that can be treated with the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein is a renal disease or disorder, a vascular disease or disorder, a skin disease or disorder, an ophthalmic disease or disorder, a nervous system disease or disorder, a blood disease or disorder, a musculoskeletal system disease or disorder, a genitourinary disease or disorder, a metabolic disease or disorder, an endocrine disease or disorder, a gastrointestinal disease or disorder, or a pulmonary disease or disorder.

[0241]

[0254] In some embodiments, the disease or disorder associated with MASP-2-dependent complement activation treatable with the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein is a kidney disease. The kidney disease may be IgA nephropathy (i.e., Berger's disease), including severe IgAN and IgAVN (IgA vasculitis-associated nephritis), lupus nephritis (LN), membranous nephropathy (MN), nephrotic syndrome, glomerular disease such as glomerulonephritis, membranous glomerulonephritis, C3 glomerulopathy (C3G), IgM nephropathy, chronic kidney disease, or chronic renal failure. In some embodiments, a method for treating IgA nephropathy in a subject in need thereof is provided, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, a method for treating severe IgAN using the anti-MASP-2 antibodies or antigen-binding fragments provided herein is provided. In some embodiments, the methods provided herein treat IgAVN using an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, a method is provided for treating lupus nephritis (LN) in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, a method is provided for treating membranous nephropathy (MN) in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, a method is provided for treating C3 glomerulopathy (C3G) in a subject in need of such treatment, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, a method is provided for treating a disease or condition associated with proteinuria in a subject in need of treatment, the method comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein.In some embodiments, a method is provided for treating a disease or condition caused by or exacerbated by fibrosis and / or inflammation in a subject in need of treatment, the method comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein.

[0242]

[0255] In some embodiments, the disease or disorder associated with MASP-2-dependent complement activation treatable with the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein is a cardiovascular disease. The cardiovascular disease can be, for example, a thrombotic disease or disorder, thrombotic microangiopathy (TMA), paroxysmal nocturnal hemoglobinuria (PNH), vasculitis, or an acquired hypercoagulable state (e.g., the presence of a catheter in a central vein). In some embodiments, a method for treating TMA in a subject in need thereof is provided, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, the disease or disorder is hematopoietic stem cell transplant-associated TMA (HSCT-TMA). In some embodiments, the disease or disorder is atypical hemolytic uremic syndrome (aHUS). In some embodiments, the disease or disorder is thrombotic thrombocytopenic purpura (TTP). In some embodiments, the disease or disorder is TMA secondary to cancer. In some embodiments, the disease or disorder is TMA secondary to chemotherapy. In some embodiments, the disease or disorder is TMA secondary to transplantation. In some embodiments, a method for treating aHUS in a subject in need of treatment is provided, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein. In some embodiments, a method for treating TTP in a subject in need of treatment is provided, comprising administering to the subject a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment provided herein.

[0243]

[0256] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein can be used to treat subjects who have undergone, are undergoing, or are about to undergo organ or tissue transplant surgery, preferably a transplant surgery selected from the group consisting of organ allotransplantation, organ xenotransplantation, and tissue transplantation. In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein can be used to treat graft-versus-host disease.

[0244]

[0257] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein can be used to treat COVID-19 complications.

[0245]

[0258] The actual dosage level of the active ingredient (i.e., anti-MASP-2 antibody or antigen-binding fragment) in the pharmaceutical compositions described herein can be varied to obtain an amount of active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without toxicity to the patient. The selected dosage level will depend on various pharmacokinetic factors, including the activity of the particular composition described herein, the route of administration, the time of administration, the rate of excretion, the duration of treatment, other drugs, compounds, and / or materials used in combination with the particular composition utilized, the age, sex, weight, condition, general health, and previous medical history of the patient being treated, and similar factors well known in the medical field.

[0246]

[0259] The anti-MASP-2 antibody or antigen-binding fragment can be administered as a sustained-release formulation, in which case less frequent administration is required. The dosage and frequency will vary depending on the half-life of the anti-MASP-2 antibody or antigen-binding fragment in the patient. In therapeutic applications, relatively high dosages at relatively short intervals may be required until disease progression slows or stops and the patient shows partial or complete improvement in disease symptoms.

[0247]

[0260] The anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein can be administered to a subject by any method known in the art, including, but not limited to, pleural, intravenous, subcutaneous, intranodal, intramuscular, intradermal, intrathecal, intrapleural, intraperitoneal, intracranial, spinal, or other parenteral routes of administration, such as by injection or infusion, or direct administration into the thymus. As used herein, the term "parenteral administration" refers to modes of administration other than enteral and topical administration, usually by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion. In some embodiments, subcutaneous administration is employed. In some embodiments, intravenous administration is employed. In some embodiments, oral administration is employed. In one embodiment, the antibodies or antigen-binding fragments provided herein can be delivered locally, hi another embodiment, the antibodies or antigen-binding fragments provided herein can be administered systemically.

[0248]

[0261] In the methods disclosed herein, a therapeutically effective amount of an anti-MASP-2 antibody or antigen-binding fragment or pharmaceutical composition disclosed herein is administered to a subject who would benefit from reduced MASP-2-dependent complement activation. The subject may be a human who would benefit from selective inhibition of lectin pathway complement activation. The subject may have undesired complement activation, unregulated complement activation, or excessive complement activation of the lectin pathway. The subject may be a mammal. In some embodiments, the subject is a human.

[0249]

[0262] The anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein can be administered using medical devices known in the art. For example, in some embodiments, needleless hypodermic injection devices can be used, such as those disclosed in U.S. Patent Nos. 5,399,163, 5,383,851, 5,312,335, 5,064,413, 4,941,880, 4,790,824, or 4,596,556. Examples of well-known implants and modules for use herein include U.S. Pat. No. 4,487,603, which discloses an implantable microinfusion pump that dispenses medication at a controlled rate; U.S. Pat. No. 4,486,194, which discloses a therapeutic device for administering drugs through the skin; U.S. Pat. No. 4,447,233, which discloses a medication infusion pump for delivering medication at a precise infusion rate; U.S. Pat. No. 4,447,224, which discloses an implantable variable flow rate infusion device for continuous drug delivery; U.S. Pat. No. 4,439,196, which discloses an osmotic drug delivery system with multiple chamber compartments; and U.S. Pat. No. 4,475,196, which discloses an osmotic drug delivery system. These patents are incorporated herein by reference. Many other such implants, delivery systems, and modules are known to those skilled in the art.

[0250]

[0263] In some embodiments, the anti-MASP-2 antibodies or antigen-binding fragments or pharmaceutical compositions provided herein can be administered in conjunction with an additional therapy. In some embodiments, the additional therapy administered in conjunction with the anti-MASP-2 antibodies or antigen-binding fragments provided herein can be an anti-inflammatory and / or analgesic, an anti-restenotic agent, or another complement inhibitor. Exemplary anti-inflammatory and / or analgesic agents include, for example, serotonin receptor antagonists; serotonin receptor agonists; histamine receptor antagonists; bradykinin receptor antagonists; kallikrein inhibitors; tachykinin receptor antagonists, including neurokinin and neurokinin receptor subtype antagonists; calcitonin gene-related peptide (CGRP) receptor antagonists; interleukin receptor antagonists; phospholipase inhibitors, including PLA2 isoform inhibitors and PLCγ isoform inhibitors, cyclooxygenase (COX) inhibitors (COX-I selective inhibitors, COX-2 selective inhibitors, or non-selective COX-I and COX-2 inhibitors), lipoxygenase inhibitors, and the like. These include inhibitors of enzymes that have therapeutic effects; prostanoid receptor antagonists, including eicosanoid EP-1 and EP-4 receptor subtype antagonists and thromboxane receptor subtype antagonists; leukotriene receptor antagonists, including leukotriene B4 receptor subtype antagonists and leukotriene D4 receptor subtype antagonists; opioid receptor agonists, including μ-opioid, δ-opioid, and κ-opioid receptor subtype agonists; purinergic receptor agonists and purinergic receptor antagonists, including P2X receptor antagonists and P2Y receptor agonists; adenosine triphosphate (ATP)-sensitive potassium channel openers; MAP kinase inhibitors; nicotinic acetylcholine inhibitors; and α-adrenergic receptor agonists. Exemplary complement inhibitors may be C3 inhibitors (e.g., pegcetacoplan) or anti-C5 antibodies (e.g., eculizumab or ravulizumab).

[0251]

[0264] The additional therapy can be administered before, simultaneously with, or after administration of the anti-MASP-2 antibodies or antigen-binding fragments, cells, or pharmaceutical compositions described herein. Combined administration can include simultaneous administration, either as a single pharmaceutical formulation or using separate formulations, or sequential administration, in any order, usually within a period of time, to allow all active agents to simultaneously exert their biological activities. Those skilled in the art can readily determine the appropriate regimen for administering the pharmaceutical compositions described herein in combination with the additional therapy, including the timing and administration of the additional agents used in the combination therapy, based on the needs of the subject being treated. 5.8 Experiments

[0265] The examples provided below are for illustrative purposes only and, unless otherwise specified, are not intended to be limiting. Accordingly, the present invention should not be construed as being limited in any way to the following examples, but rather as embracing any and all variations that become evident as a result of the teachings provided herein.

[0252]

[0266] Briefly, the results of the studies described below show that clone 3E10 and its humanized variants bind to the protease domain of human MASP-2 with high affinity and are far more effective than the benchmark antibody narsoplimab ("MASP-2-BM") in inhibiting complement activation. 5.8.1 Example 1: Immunization and Antibody Screening

[0267] Mice from various strains (C57BL / 6, BALB / c, SJL, and CD-1) were immunized with recombinant human MASP-2 protein (SEQ ID NO: 1, NCBI accession number NP_006601.2). Those that generated strong titer responses were selected for single B cell isolation. B cells from the spleen and lymph nodes were isolated and enriched with microbeads. B cells that recognized human MASP-2 were stained and isolated by FACS. The immunoglobulin heavy and light chain gene sequences of these B cells were cloned and recombinantly expressed. These monoclonal antibodies were then rescreened for their binding and blocking activity against human MASP-2. Clone 3E10 was selected for further study. 5.8.2 Example 2: Evaluation of Blocking Activity Using a C4 Activation Assay

[0268] Methods: The blocking activity of chimeric antibodies was evaluated using a C4 activation assay. 100 μl of 10 μg / mL mannan (Sigma-M7504) was coated onto a 96-well ELISA plate overnight at 4°C. The antibody was serially diluted in C4 activation buffer (0.1% BSA + 20 mM Tris-HCl + 2 mM CaCl2 + 140 mM NaCl + 1 mM MgCl2 + 0.05% Tween 20, pH 7.4). 55 μl of the serially diluted antibody was mixed with 55 μl of C4 activation buffer containing 2% human serum (Quidel-A113) and incubated on ice for 45 minutes. The mannan-coated plate was washed with washing buffer (10 mM Tris-HCl + 140 mM NaCl + 0.05% Tween 20 + 2 mM CaCl2, pH 7.2-7.4) and blocked with blocking buffer (0.1% BSA + 10 mM Tris-HCl + 140 mM NaCl, pH 7.4). 100 μl of the antibody-serum mixture was added to the mannan-coated plate and incubated at 37°C for 1.5 hours. After washing, 100 μl of HRP-linked anti-C4 antibody (Quidel-A211, 1:3000) was added to the plate and incubated for 1 hour. TMB was used to detect activated complement C4.

[0253]

[0269] Results and Conclusions: As shown in Figures 1 and 2, the chimeric antibodies inhibited C4 activation to different degrees. Clone 3E10 was the most effective, with its IC 50 The IC of MASP-2-BM was 0.003603 μg / mL. 50 was 0.2416 μg / mL. 5.8.3 Example 4: Inhibition of C3 and MAC Activation by 3E10

[0270] Methods: C3 and MAC activation assay: 100 μl of 10 μg / mL mannan (Sigma-M7504) was coated onto a 96-well ELISA plate overnight at 4°C. Antibodies were serially diluted in activation buffer (0.1% BSA + 20 mM Tris-HCl + 2 mM CaCl2 + 140 mM NaCl + 1 mM MgCl2 + 0.05% Tween 20, pH 7.4). 55 μl of serially diluted antibodies were mixed with 55 μl of activation buffer containing 2% human serum (Quidel-A113) and incubated on ice for 45 minutes. Mannan-coated plates were washed with wash buffer (10 mM Tris-HCl + 140 mM NaCl + 0.05% Tween 20 + 2 mM CaCl2, pH 7.2–7.4) and blocked with blocking buffer (0.1% BSA + 10 mM Tris-HCl + 140 mM NaCl, pH 7.4). 100 μl of the antibody-serum mixture was added to the mannan-coated plates and incubated for 1.5 h at 37°C. After washing, 100 μl of HRP-linked anti-C3 antibody (Quidel-A205, 1:3000) or HRP-linked anti-MAC antibody (Quidel-A239, 1:3000) was added to the plates, respectively, and incubated for 1 h. Activated complement C3 or MAC was detected using TMB (3,3',5,5''-tetramethylbenzidine).

[0254]

[0271] Results and Conclusions: As shown in Figures 4 and 5, chimeric 3E10 had an IC of 0.004142 μg / mL. 50 inhibited C3 activation with an IC of 0.003266 μg / mL 50 inhibited MAC activation. 5.8.4 Example 3: Binding of 3E10 to MASP-2 as measured by BLI

[0272] Methods: Clone 3E10 was diluted to 100 nM in kinetics buffer (PBS pH 7.4, 0.1% BSA + 0.01% Tween-20). Recombinant MASP-2 protein was diluted in kinetics buffer to obtain a concentration gradient: 200 nM, 100 nM, 50 nM, 25 nM, and 0 nM (reference control). The antibody was immobilized on a protein A biosensor after equilibration. The baseline was detected for 60 seconds. Subsequently, antibody-antigen binding was detected for 120 seconds, and the K on Then, dissociation was detected in the kinetic buffer for 120 seconds, and K off Data were obtained. Regeneration of the biosensor was performed in 10 mM glycine, pH 2.0 buffer. All kinetic data were collected at 30°C. Data were acquired on a Gator Bioanalysis System.

[0255]

[0273] Results and Conclusions: As shown in Figure 3 and Table 4, chimeric 3E10 exhibited high affinity (K D It bound to human MASP-2 with a binding affinity of <1E-10 M.

[0256] [Table 5]

[0257] 5.8.5 Example 5: Inhibition of C4 activation in human serum at different concentrations by 3E10

[0274] Methods: The blocking activity of 3E10 in different concentrations of human serum was evaluated using a C4 activation assay. 100 μl of 10 μg / mL mannan (Sigma-M7504) was coated onto a 96-well ELISA plate overnight at 4°C. The antibody was serially diluted in C4 activation buffer (0.1% BSA + 20 mM Tris-HCl + 2 mM CaCl2 + 140 mM NaCl + 1 mM MgCl2 + 0.05% Tween 20, pH 7.4). 55 μl of the serially diluted antibody was mixed with 55 μl of C4 activation buffer containing different concentrations of human serum (Quidel-A113) and incubated on ice for 45 minutes. The final concentrations of human serum were 5%, 25%, and 50%, respectively. The mannan-coated plate was washed with wash buffer (10 mM Tris-HCl + 140 mM NaCl + 0.05% Tween 20 + 2 mM CaCl2, pH 7.2-7.4) and blocked with blocking buffer (0.1% BSA + 10 mM Tris-HCl + 140 mM NaCl, pH 7.4). 100 μl of the antibody-serum mixture was added to the mannan-coated plate and incubated at 37°C for 1.5 hours. After washing, 100 μl of HRP-linked anti-C4 antibody (Quidel-A211, 1:3000) was added to the plate and incubated for 1 hour. TMB was used to detect activated complement C4.

[0258]

[0275] Results and Conclusions: As shown in Figure 6, chimeric 3E10 exhibited IC values of 0.008259 μg / mL, 0.03085 μg / mL, and 0.06734 μg / mL in 5%, 10%, and 50% human serum, respectively. 50 inhibited the activation of C4. 5.8.6 Example 6: Humanization of 3E10 and Binding Affinity of 3E10 to MASP-2 as Measured by BLI

[0276] Methods: For the humanization of 3E10, IgBLAST from NCBI was used to select the optimal human framework for rodent CDR grafting. Variable regions with high amino acid sequence identity (homologous match or best match) with the rodent variable regions were used. 3E10 was humanized by grafting three CDRs from the light chain variable region into a human VL that was as homologous as possible to the mouse antibody VL. Similarly, the three CDRs from the heavy chain variable region were grafted into a human VH that was as homologous as possible to the mouse antibody VL. Kabat numbering was then performed. Furthermore, several amino acid residues in the framework regions of the selected human variable regions were changed to amino acid residues present in the mouse variable region (called "backmutation"). The humanized sequence of 3E10 is shown in Table 3 above. Different humanized clones share a single light chain sequence. The binding and activity of the humanized 3E10 clones were evaluated by BLI and C4 activation assays. The experimental procedures for the C4 activation assay were similar to those described in the previous examples.

[0259]

[0277] The binding affinity of the humanized 3E10 clone was measured by BLI using a gator. The humanized 3E10 clone or MSAP-2-BM was immobilized on a Protein A biosensor. The assay was performed at 30°C in 1x PBS (136 mM NaCl, 8 mM NaHPO4, 2 mM KH2PO4, 2.6 mM KCl) containing 0.1% BSA and 0.005% Tween-20, pH 7.4. Diluted antibody and human MASP-2 were added to the corresponding wells. After loading of human MASP-2, the association and dissociation of the antibody were detected by the biosensor. The biosensor was regenerated in 10 mM glycine, pH 2.0 buffer.

[0260]

[0278] Results and Conclusions: As shown in Table 5, the humanized 3E10 clone maintained high binding affinity for human MASP-2, comparable to that of chimeric 3E10. Furthermore, as shown in Figure 7, humanized 3E10 exhibited high C4 blocking activity, comparable to that of chimeric 3E10.

[0261] [Table 6]

[0262] 5.8.7 Example 7: Binding of humanized 3E10 to MASP-2 measured by SPR

[0279] Methods: The binding affinity of humanized 3E10 was further measured by surface plasmon resonance (SPR) technology using a Biacore 8K. Humanized 3E10 or MASP-2-BM was immobilized on a CM-5 chip. The assay was performed at 25°C, and the running buffer was 1x HEPES (10 mM HEPES, 150 mM NaCl, 3 mM EDTA) containing 0.005% Tween-20 and pH 7.4. Diluted antibodies were captured on the sensor chip using the Fc capture method. Human MASP-2 was used as the analyte, and the running buffer was injected as the dissociation phase.

[0263]

[0280] Results and Conclusions: Representative results are shown in Figure 8 and Table 6, demonstrating that humanized 3E10 has a much lower K than the benchmark antibody. D We confirmed that it binds to MASP-2.

[0264] [Table 7]

[0265] 5.8.8 Example 8: Inhibition of C4, C3, and MAC Activation by Humanized 3E10

[0281] Methods: Human serum samples were collected from 10 healthy donors by standard methods without the use of clot activators. C4, C3, and MAC assays were performed using procedures similar to those described in the previous examples for the humanized 3E10 disclosed herein.

[0266]

[0282] Results and Conclusions: Representative results are shown in Figure 9. As shown, humanized 3E10 had a much lower IC than the benchmark antibody. 50IC values of 0.006200 μg / mL, 0.02232 μg / mL, and 0.009792 μg / mL, respectively. 50 At these values, it strongly inhibited the activation of C4, C3, and MAC in serum from healthy donors. 5.8.9 Example 9: Humanized 3E10 did not affect either the classical or alternative pathway

[0283] Methods: For classical pathway activation, 3 μg / mL IgM antibody was coated onto 96-well ELISA plates overnight at 4°C. The antibody was serially diluted in activation buffer (0.1% BSA + 20 mM Tris-HCl + 2 mM CaCl2 + 140 mM NaCl + 1 mM MgCl2 + 0.05% Tween 20, pH 7.4) containing 40% human serum, mixed, and incubated on ice for 45 minutes. 10 mM EDTA was used as a positive control. The IgM-coated plate was washed with wash buffer (10 mM Tris-HCl + 140 mM NaCl + 0.05% Tween 20 + 2 mM CaCl2, pH 7.2–7.4) and blocked with blocking buffer (0.1% BSA + 10 mM Tris-HCl + 140 mM NaCl, pH 7.4). 100 μl of the antibody-serum mixture was added to the IgM-coated plate and incubated for 1.5 hours at 37°C. After washing, 100 μl of HRP-linked anti-MAC antibody was added to the plate and incubated for 1 hour. TMB was used to detect MAC formation.

[0267]

[0284] For activation of the alternative pathway, 3 μg / mL LPS was diluted in PBS containing 10 mM MgCl2 and coated onto a 96-well ELISA plate overnight at 4°C. The antibody was serially diluted in activation buffer (20 mM Tris-HCl + 10 mM EGTA + 5 mM MgCl2, pH 7.4) containing 10% human serum, mixed, and incubated on ice for 45 minutes. 10 mM EDTA was used as a positive control. The LPS-coated plate was washed with wash buffer (10 mM Tris-HCl + 140 mM NaCl + 0.05% Tween 20 + 2 mM CaCl2, pH 7.2–7.4). 100 μl of the antibody-serum mixture was added to the LPS-coated plate and incubated for 1.5 hours at 37°C. After washing, 100 μl of HRP-linked anti-MAC antibody was added to the plate and incubated for 1 hour. TMB was used to detect MAC formation.

[0268]

[0285] Results and Conclusions: Representative results are shown in Figure 10. As shown, humanized 3E10 did not affect MAC deposition through activation of either the classical or alternative pathways. 5.8.10 Example 10: Epitope Binning of 3E10

[0286] Methods: MASP-2-his, 3E10, or MASP-2-BM was diluted to a concentration of 100 nM in kinetic buffer (PBS pH 7.4, 0.1% BSA + 0.01% Tween-20). MASP-2-his was immobilized on an anti-his biosensor after equilibration. Baseline detection was performed for 60 seconds. Binding of the first antibody was detected for 240 seconds, followed by equilibration. Subsequently, binding of the second antibody was detected for 240 seconds. Biosensor regeneration was performed in 10 mM glycine, pH 2.0 buffer. All kinetic data were collected at 30°C. Data were acquired using a Gator Bioanalysis System.

[0269]

[0287] Results and Conclusions: As shown in Figure 11, the binding of the first antibody did not affect the binding of the second antibody, indicating that 3E10 and MASP-2-BM bind to different epitopes on MASP-2. 5.8.11 Example 11: Epitope Binning of Humanized 3E10

[0288] Methods: As shown in Figure 12, truncated human MASP-2 proteins were designed for epitope mapping. The sequences of these antigens are shown in Table 7. Genes encoding these antigens were synthesized and expressed in a CHO expression system. Binding of the humanized 3E10 or benchmark antibodies disclosed herein to different truncated human MASP-2 proteins was tested by ELISA.

[0270] [Table 8-1]

[0271] [Table 8-2]

[0272]

[0289] Results and Conclusions: Representative results are shown in Figure 13. As shown, humanized 3E10 completely lost binding to truncated human MASP-2 lacking the protease domain, indicating that the protease domain of human MASP-2 is required for binding of humanized 3E10 to truncated human MASP-2. In contrast, the benchmark antibody MASP-2-BM showed different binding properties for the truncated protein and retained some binding to human MASP-2 lacking the protease domain, confirming that humanized 3E10 and MASP-2-BM bind to different epitopes on MASP-2. 6. Reference to Electronically Submitted Sequence Listings This application incorporates by reference the sequence listing that accompanies this application as an xml file entitled "[P23400663C].SEQ.xml", created on March 18, 2022, and having a size of 204,816 bytes.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to human MASP-2, (1) As defined by Kabat: (a) a light chain variable region (VL) comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9, and 11, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VL CDRs; and / or (b) a heavy chain variable region (VH) comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 12, 16, and 18, respectively, or a variant thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDRs; or (2) When defined by IMGT, (a) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 8, 10, and 11, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VL CDRs; and / or (b) An antibody or antigen-binding fragment thereof comprising a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 13, 17, and 19, respectively, or variants thereof having up to about 5 amino acid substitutions, additions, and / or deletions in the VH CDRs.

2. 2. The antibody or antigen-binding fragment of claim 1, comprising a VL CDR1, a VL CDR2, a VL CDR3, a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 7, 9, 11, 12, 16, and 18, respectively, as defined by Kabat.

3. 2. The antibody or antigen-binding fragment of claim 1, wherein, as defined by IMGT, the antibody or antigen-binding fragment comprises a VL CDR1 having the amino acid sequence of SEQ ID NO: 8, a VL CDR2 having the amino acid sequence of SEQ ID NO: 10, a VL CDR3 having the amino acid sequence of SEQ ID NO: 11, a VH CDR1 having the amino acid sequence of SEQ ID NO: 13, 14 or 15, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 19 or 20.

4. (1) SEQ ID NOs: 8, 10, 11, 13, 17 and 19, respectively; (2) SEQ ID NOs: 8, 10, 11, 14, 17, and 19, respectively; (3) SEQ ID NOs: 8, 10, 11, 15, 17, and 19, respectively; (4) SEQ ID NOs: 8, 10, 11, 13, 17 and 20, respectively; (5) SEQ ID NOs: 8, 10, 11, 14, 17, and 20, respectively; or (6) SEQ ID NOs: 8, 10, 11, 15, 17 and 20, respectively 4. The antibody or antigen-binding fragment of claim 3, comprising a VL CDR1, a VL CDR2, a VL CDR3, a VH CDR1, a VH CDR2 and a VH CDR3 having the amino acid sequences:

5. An antibody or antigen-binding fragment thereof that specifically binds to human MASP-2, (a) a VL having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 21; and / or (b) an antibody or antigen-binding fragment thereof comprising a VH having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:

22.

6. The antibody or antigen-binding fragment of claim 5, comprising a VL having the amino acid sequence of SEQ ID NO: 21 and a VH having the amino acid sequence of SEQ ID NO:

22.

7. An antibody or antigen-binding fragment thereof that specifically binds to human MASP-2, (a) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 derived from a VL having the amino acid sequence of SEQ ID NO: 21; and / or (b) an antibody or antigen-binding fragment thereof, comprising a VH comprising VH CDR1, VH CDR2 and VH CDR3 derived from the VH having the amino acid sequence of SEQ ID NO:

22.

8. 8. The antibody or antigen-binding fragment of claim 1 , which is a chimeric antibody or antigen-binding fragment, a humanized antibody or antigen-binding fragment, or a human antibody or antigen-binding fragment.

9. 9. The antibody or antigen-binding fragment of claim 8, which is a humanized antibody or antigen-binding fragment.

10. (a) a VL having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 23, and / or (b) the antibody or antigen-binding fragment of claim 9, comprising a VH having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:

24.

11. The antibody or antigen-binding fragment of claim 10, comprising a VL having the amino acid sequence of SEQ ID NO: 23 and a VH having an amino acid sequence selected from the group consisting of SEQ ID NOs: 24 to 32.

12. Fab, Fab', F(ab') ２ , Fv, scFv, (scFv) ２ 12. The antibody or antigen-binding fragment of any one of claims 1 to 11, selected from the group consisting of: a single domain antibody (sdAb) and a heavy chain antibody (HCAb).

13. The antibody or antigen-binding fragment of any one of claims 1 to 11, which is an IgG1 antibody or variant thereof, an IgG2 antibody or variant thereof, an IgG3 antibody or variant thereof, or an IgG4 antibody or variant thereof.

14. The antibody of claim 13, which is an IgG4 antibody or a variant thereof.

15. (1) a light chain having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 33; (2) The antibody described in claim 14, comprising a heavy chain having at least 85%, at least 90%, at least 95%, at least 98%, or 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO:

34.

16. 16. The antibody of claim 15, wherein the heavy chain has an amino acid sequence selected from the group consisting of SEQ ID NOs: 34-42, or a variant thereof modified by one or more amino acid substitutions that increase the terminal half-life of the antibody.

17. 17. The antibody of claim 16, wherein the heavy chain variant is modified by one or more substitutions at amino acid residues selected from the group consisting of S228, F234, L235, M252, S254, T256, K288, T307, M428, N434, H435 and Y436 (numbered according to the EU index).

18. The heavy chain variants are selected from the group consisting of: i) S228P, ii) F234A and L235A, iii) S228P, F234A and L235A, iv) T307H and N434A, v) M252Y, S254T and T256E, vi) M428L, N434A and Y436T, vii) S228P, M252Y, S254T and T256E, vi 17. The antibody of claim 16, wherein the antibody is modified by amino acid substitutions selected from the group consisting of: (i) S228P, F234A, L235A, M252Y, S254T and T256E; (ix) S228P, F234A, L235A, T307Q and N434A; and (x) M252Y, S254T, T307H and N434A.

19. The antibody of claim 15, wherein the light chain has the amino acid sequence of SEQ ID NO: 33 and the heavy chain has an amino acid sequence selected from the group consisting of SEQ ID NOs: 43-74.

20. 20. An antibody or antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment of any one of claims 1 to 19 for binding to human MASP-2.

21. 21. The antibody or antigen-binding fragment of any one of claims 1 to 20, which is a bispecific or multispecific antibody.

22. 22. The antibody or antigen-binding fragment of any one of claims 1 to 21, which is a monoclonal antibody or antigen-binding fragment.

23. (1) a K of about 1 nM or less as measured by surface plasmon resonance (SPR) Ｄ binds to human MASP-2 at (2) inhibits the activation of C4 with an IC50 of about 0.1 μg / mL or less as measured in vitro; (3) inhibits C3 activation with an IC50 of about 0.1 μg / mL or less as measured in vitro; (4) inhibiting MAC activation with an IC50 of about 0.1 μg / mL or less as measured in vitro; (5) does not affect the classical or alternative pathways of complement activation; or (6) Having any combination of the properties of (1) to (5), 23. The antibody or antigen-binding fragment of any one of claims 1 to 22.

24. An antibody or antigen-binding fragment thereof that specifically binds to the protease domain of human MASP-2, (1) K<1 nM as measured by SPR Ｄ binds to human MASP-2 at (2) inhibits C4 activation with an IC50 of 0.1 μg / mL or less as measured in vitro; (3) inhibits C3 activation with an IC50 of 0.1 μg / mL or less as measured in vitro; (4) inhibiting MAC activation with an IC50 of 0.1 μg / mL or less as measured in vitro; (5) does not affect the classical or alternative pathways of complement activation; or (6) Having any combination of the properties of (1) to (5), An antibody or antigen-binding fragment thereof.

25. 25. The antibody or antigen-binding fragment of claim 23 or 24, which inhibits C4 activation with an IC50 of 0.001 to 0.01 μg / mL measured in vitro.

26. 25. The antibody or antigen-binding fragment of claim 23 or 24, which inhibits C4 activation in the presence of 5-50% human serum with an IC50 of 0.1 μg / mL or less as measured in vitro.

27. K in the range of 0.01 nM to 1 nM as measured by SPR Ｄ 27. The antibody or antigen-binding fragment of any one of claims 23 to 26, which binds to human MASP-2 at

28. K in the range of 0.05 nM to 0.5 nM as measured by SPR Ｄ 28. The antibody or antigen-binding fragment of claim 27, which binds to human MASP-2 at

29. 29. A polynucleotide encoding the antibody or antigen-binding fragment of any one of claims 1 to 28.

30. A vector comprising the polynucleotide of claim 29.

31. 31. A host cell comprising the polynucleotide of claim 29 or the vector of claim 30.

32. 30. A pharmaceutical composition comprising a therapeutically effective amount of the antibody or antigen-binding fragment of any one of claims 1 to 28 and a pharmaceutically acceptable carrier.

33. 30. A method of inhibiting MASP-2-dependent complement activation in a subject in need thereof, comprising administering to the subject an effective amount of the antibody or antigen-binding fragment of any one of claims 1 to 28.

34. 34. The method of claim 33, wherein the subject has a disease or disorder associated with MASP-2-dependent complement activation.

35. 29. A method of treating a disease or disorder associated with MASP-2-dependent complement activation in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment of any one of claims 1 to 28.

36. 36. The method of claim 35, further comprising administering to the subject an additional therapy.

37. 37. The method of any one of claims 33 to 36, wherein the subject is a human.

38. 29. Use of the antibody or antigen-binding fragment of any one of claims 1 to 28 in the treatment of a disease or disorder associated with MASP-2-dependent complement activation.

39. 30. Use of the antibody or antigen-binding fragment of any one of claims 1 to 28 for the preparation of a medicament for the treatment of a disease or disorder associated with MASP-2-dependent complement activation.

40. 40. The method or use of any one of claims 34 to 39, wherein the disease or disorder is a kidney disease or disorder, a vascular disease or disorder, a skin disease or disorder, an ophthalmological disease or disorder, a nervous system disease or disorder, a blood disease or disorder, a musculoskeletal disease or disorder, a genitourinary disease or disorder, a metabolic disease or disorder, an endocrine disease or disorder, a gastrointestinal disease or disorder, or a pulmonary disease or disorder.

41. 40. The method or use of any one of claims 34 to 39, wherein the disease or disorder is IgA nephropathy (IgAN), thrombotic microangiopathy (TMA), lupus nephritis (LN), membranous nephropathy (MN), or C3 glomerulopathy (C3G).

42. 42. The method or use of claim 41, wherein the disease or disorder is IgAN.

43. 42. The method or use of claim 41, wherein the disease or disorder is TMA.

44. 44. The method or use of claim 43, wherein the disease or disorder is atypical hemolytic uremic syndrome (aHUS), hematopoietic stem cell transplant-associated TMA (HSCT-TMA), thrombotic thrombocytopenic purpura (TTP), TMA secondary to cancer, TMA secondary to chemotherapy, or TMA secondary to transplantation.

45. 45. The method or use of claim 44, wherein the disease or disorder is HSCT-TMA.

46. 45. The method or use of claim 44, wherein the disease or disorder is TTP.

Images