Development and application of a complement inhibitor

Abstract

The present disclosure relates to the development and application of a complement inhibitor, which is an antibody or an antigen-binding portion thereof that binds to MASP-2. The antibody disclosed herein can bind to MASP-2 with high affinity, can specifically recognize the enzymatic domains CCP1-CCP2-SP of human, monkey, mouse and rat MASP-2, and can effectively inhibit the generation of complement component C4b and C3 convertase C4b2b under low concentration conditions, and its blocking activity is 50-200 times higher than that of known antibodies, and the MASP-2 antibody is well tolerated without any toxic reactions, and its efficacy and safety are better than similar antibodies.

Description

Technical Field

[0001] The present disclosure relates to a complement inhibitor and its application, in particular to a MASP-2 inhibitory antibody and its application. Background Art

[0002] Mannose-binding lectin-associated serine protease 2 (MASP-2) belongs to the serine protease family and is a key enzyme in the complement lectin activation pathway. MASP-2 contains six domains, namely N-terminal CUB1, EGF, CUB2, CCP1, CCP2 and C-terminal SP enzymatic domain, with a molecular weight of about 75kDa. MASP-2 in the blood binds to the lectin pathway recognition molecules mannose-binding lectin (MBL), collectin 11 (CL-K1) and ficolin (ficolin-1, ficolin-2 and ficolin-3) through the three N-terminal domains of CUB1-EGF-CUB2 to form a complex. When MBL, CL-K1 or ficolin binds to pathogen-associated molecular patterns (PAMPs) such as D-mannose, N-acetyl-D-glucosamine or acetyl, MASP-1 and MASP-2 are activated sequentially. The activated MASP-2 cleaves C4 and C2. The cleaved fragment C4b interacts with C2b to produce C3 convertase (C4b2b). C3 convertase (C4b2b) activates C3, leading to the production of C5 convertase (C4b2b3b) and the formation of membrane attack complex (C5b-9) that can cause microbial lysis. The activated forms of C3 and C4 (C3b and C4b) are covalently deposited on the surface of foreign targets, which are recognized by complement receptors on a variety of phagocytes and phagocytize the corresponding pathogens. The lectin pathway plays an important role in the body's natural immunity against pathogenic microorganisms such as bacteria, yeast, and viruses.

[0003] Under certain pathological conditions, after cells are damaged and ruptured, the incompletely glycosylated glycoproteins (containing high-mannose "precursor" polysaccharide clusters) inside them are released, which can be recognized by MBL with high affinity and initiate complement activation (Maynard et al., J. Biol. Chem. 257: 3788-3794, 1982); similarly, immune complexes formed by IgA antibodies with galactose modification defects are deposited on the inner wall of glomerular blood vessels, etc., and the complement lectin pathway is activated through MBL-MASP-2, causing an inflammatory response. Granzymes released by overactivated inflammatory cells may cause damage to nearby tissues and organs. In addition, activated complement components are deposited on nearby cells, which can lead to the production of complement membrane attack complexes and cell lysis. The complement system is associated with the pathogenesis of a variety of acute or chronic disease conditions, such as IgA nephropathy (IgAN), ischemia-reperfusion injury, transplant rejection, rheumatoid arthritis, myocardial infarction, vascular remodeling after stroke, ARDS, septic shock, capillary leakage after thermal burns, inflammation after cardiopulmonary bypass, multiple sclerosis, myasthenia gravis, and Alzheimer's disease. Recent studies have shown that multiple tissue damage such as lungs and kidneys caused by the new coronavirus COVID-2019 infection is related to the complement lectin pathway (Magro C et al. Transl Res. 2020 Jun; 220: 1-13), and inhibiting MASP-2 activity can greatly reduce the mortality of severe COVID-2019 patients (Rambaldi A et al. Immunobiology. 2020 Aug 9: 152001).

[0004] MASP-2 is the only protease in the lectin pathway that can cleave C4. Its concentration in healthy human blood is much lower than that of MASP-1 and other complement components in the lectin pathway. Kidmose R et al. (PNAS September 18, 2012 109(38)15425-15430) showed that MASP-2 binds to C4 through the CCP2 domain, causing the latter to undergo conformational changes and expose the cleavage site. Mutations in the CCP2 domain can completely inhibit the activity of MASP-2. Summary of the invention

[0005] The present disclosure relates to anti-MASP-2 inhibitory antibodies for use in inhibiting the adverse effects of MASP-2-dependent complement activation.

[0006] In one aspect, the disclosure provides an antibody, or antigen-binding portion thereof, that binds MASP-2.

[0007] In one aspect, the disclosure provides a bispecific antibody, or an antigen-binding portion thereof.

[0008] In one aspect, the disclosure provides nucleic acids encoding the aforementioned antibodies, or antigen-binding portions thereof, or bispecific antibodies, or antigen-binding portions thereof, that bind MASP-2.

[0009] In one aspect, the present disclosure provides a vector comprising the aforementioned nucleic acid.

[0010] In one aspect, the present disclosure provides a cell comprising the aforementioned vector.

[0011] In one aspect, the present disclosure provides an antibody or antigen-binding portion thereof, encoding nucleic acid, vector, cell, composition or kit comprising the aforementioned.

[0012] In one aspect, the present disclosure provides a method of treating a disease associated with MASP-2-dependent complement activation, comprising the step of administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof, nucleic acid, vector, cell and / or composition of any of the preceding aspects.

[0013] In one aspect, the disclosure provides use of an antibody or antigen-binding fragment thereof, nucleic acid, vector, cell and / or pharmaceutical composition of any of the foregoing aspects in the preparation of a medicament or kit for treating a disease associated with MASP-2-dependent complement activation in a subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Shown are the SDS-PAGE results of MASP-2D4-6 recombinant protein.

[0015] Figure 2 Inhibition of C4b deposition by hybridoma supernatants is shown.

[0016] Figure 3 Inhibition of C4b deposition by hybridoma clone chimeric antibodies is shown.

[0017] Figure 4 Inhibition of C3b deposition by hybridoma clone chimeric antibodies is shown.

[0018] Figure 5 ELISA testing of the binding of candidate chimeric antibodies to MASP-2D4-6 of different species is shown.

[0019] Figure 6 Inhibition of C4b deposition by phage display library Fab candidate clones is shown.

[0020] Figure 7 Shown is the inhibition of C4b deposition after purification of Fab candidate clones from the phage display library.

[0021] Figure 8The inhibition of C4b deposition after mutation of 28O14 clone is shown. Among them, Ser102 represents wild-type antibody, and its position 102 is serine. Gly102, Tyr102, Ala102, Glu102, Gln102, Val102, Leu102 and His102 represent mutant antibodies, and its position 102 is mutated from serine to glycine, tyrosine, alanine, glutamic acid, glutamine, valine, leucine and histidine respectively.

[0022] Figure 9A , B shows inhibition of C4b deposition by humanized MASP-2 antibodies.

[0023] Figure 10A , B, C show the inhibition of C3b deposition by humanized MASP-2 antibodies.

[0024] Figure 11 ELISA testing of binding of humanized MASP-2 antibodies to antigens from different species is shown.

[0025] Figure 12 The efficacy of humanized MASP-2 antibodies in a mouse hemolytic uremic syndrome model is shown. DETAILED DESCRIPTION

[0026] I. Definitions

[0027] In the present disclosure, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the protein and nucleic acid chemistry, molecular biology, cell and tissue culture, microbiology, immunology and laboratory operation procedures used herein are terms and routine procedures widely used in the corresponding fields. At the same time, in order to better understand the present disclosure, the definitions and explanations of the relevant terms are provided below.

[0028] Provided herein are antibodies (e.g., monoclonal antibodies) that specifically bind to MASP-2 and antigen-binding fragments thereof. In specific aspects, provided herein are monoclonal anti-MASP-2 antibodies that specifically bind to MASP-2, wherein the anti-MASP-2 antibodies comprise variants of a parent antibody. In specific aspects, provided herein are antibodies that specifically bind to MASP-2 (e.g., human MASP-2). In particular aspects, provided herein are anti-MASP-2 antibodies comprising a modification in one or more amino acid residues (e.g., 5-13 amino acid substitutions in the framework region of the heavy chain variable region) that retains affinity for the antigen compared to the parent antibody without the modification. The term "MASP-2" refers to any MASP-2 molecule known to those skilled in the art. For example, the MASP-2 can be from a mammal, for example, the MASP-2 can be from a human.

[0029] As used herein, the term "MASP-2-dependent complement activation" includes MASP-2-dependent activation of the lectin pathway, which occurs under physiological conditions (i.e., in the presence of Ca 2+ ), leading to the formation of the lectin pathway C3 convertase C4b2a, and following the accumulation of the C3 cleavage product C3b, leading to the C5 convertase C42a(C3b)n.

[0030] As used herein, the term "MASP-2 inhibitory antibody" refers to any anti-MASP-2 antibody or MASP-2 binding fragment thereof that binds to or directly interacts with MASP-2 and effectively inhibits MASP-2-dependent complement activation. The MASP-2 inhibitory antibodies used in the methods of the present disclosure can reduce MASP-2-dependent complement activation by greater than 20%, such as greater than 30%, or greater than 40%, or greater than 50%, or greater than 60%, or greater than 70%, or greater than 80%, or greater than 90%, or greater than 95%.

[0031] The term "lectin pathway" refers to complement activation that occurs through the specific binding of serum and non-serum carbohydrate binding proteins, including mannan-binding lectin (MBL) and ficolins.

[0032] As used herein and unless otherwise specified, the term "about" or "approximately" means within plus or minus 10% of a given value or range. Where an integer is required, the term means within plus or minus 10% of a given value or range, rounded up or down to the nearest integer.

[0033] With respect to antibody chain polypeptide sequences, the phrase "substantially identical" may be understood as antibody chains that exhibit at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity to a reference polypeptide sequence. With respect to nucleic acid sequences, the term may be understood as nucleotide sequences that exhibit at least greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity to a reference nucleic acid sequence.

[0034] Sequence "identity" or "identity" has a meaning recognized in the art, and the percentage of sequence identity between two nucleic acid or polypeptide molecules or regions can be calculated using published techniques. Sequence identity can be measured along the full length of a polynucleotide or polypeptide or along a region of the molecule (see, e.g., Computational Molecular Biology, Lesk, AM, ed., Oxford University Pres, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, AM, and Griffin, HG, eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991). Although there are many methods to measure the identity between two polynucleotides or polypeptides, the term "identity" is well known to those of skill (Carrillo, H. & Lipman, D., SIAM J Applied Math 48: 1073 (1988)).

[0035] A "substitution" variant is a variant in which at least one amino acid residue in the native sequence is removed and a different amino acid is inserted in the same position. The substitution may be single, in which only one amino acid is substituted in the molecule, or multiple, in which two or more amino acids are substituted in the same molecule. Multiple substitutions may be located at consecutive sites. Likewise, an amino acid may be substituted by multiple residues, in which such variants include both substitutions and insertions. An "insertion" variant is a variant in which one or more amino acids are inserted into an amino acid immediately adjacent to a particular position in a native sequence. Immediately adjacent to an amino acid means connected to the α-carboxyl or α-amino functional group of the amino acid. A "deletion" variant is a variant in which one or more amino acids in the native amino acid sequence are removed. Typically, a deletion variant has one or two amino acids deleted in a specific region of the molecule.

[0036] With respect to the variable domains of antibodies, the term "variable" refers to certain parts of related molecules that have extensive sequence differences between antibodies and are used for specific recognition and binding of a particular antibody to its specific target. However, the variability is not evenly distributed throughout the variable domain of an antibody. The variability is concentrated in three segments called complementarity determining regions (CDRs; i.e., CDR1, CDR2, and CDR3) or hypervariable regions, which are located in the variable domains of both the light and heavy chains. The more conserved portions of the variable domains are called framework (FR) regions or framework sequences. Each variable domain of a native heavy and light chain includes four FR regions, which primarily adopt a β-sheet configuration, connected by three CDRs, which form loops that connect the β-sheet structure and in some cases form part of the β-sheet structure. The CDRs of each chain are usually linked together in proximity by the FR regions and, with the help of the CDRs from the other chain, contribute to the formation of the antibody target binding site (epitope or determinant) (see Kabat et al. Sequences of Proteins of Immunological Interest, National Institute of Health, Bethesda, MD (1987)). As used herein, the numbering of immunoglobulin amino acid residues is based on the immunoglobulin amino acid residue numbering system of Kabat et al., unless otherwise indicated. A CDR may have the ability to specifically bind to a cognate epitope.

[0037] As used herein, an "antibody fragment" or "antigen-binding fragment" of an antibody refers to any portion of a full-length antibody that is less than the full length but contains at least a portion of the variable region of the antibody that binds to an antigen (e.g., one or more CDRs and / or one or more antibody binding sites), and thus retains binding specificity and at least a portion of the specific binding ability of the full-length antibody. Thus, an antigen-binding fragment refers to an antibody fragment that contains an antigen-binding portion that binds to the same antigen as the antibody from which the antibody fragment was derived. Antibody fragments include antibody derivatives produced by enzymatic treatment of full-length antibodies, as well as synthetically produced derivatives, such as recombinantly produced derivatives. Antibodies include antibody fragments. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab'), 2, single-chain Fv (scFv), Fv, dsFv, diabodies, Fd and Fd' fragments and other fragments, including modified fragments (see, e.g., Methods in Molecular Biology, Vol 207: Recombinant Antibodies for Cancer Therapy Methods and Protocols (2003); Chapter 1; p 3-25, Kipriyanov). The fragment may include multiple chains linked together, for example, by disulfide bonds and / or by peptide linkers. Antibody fragments generally contain at least or about 50 amino acids, and typically at least or about 200 amino acids. Antigen-binding fragments include any antibody fragment that, when inserted into an antibody framework (e.g., by replacing the corresponding region), obtains immunospecific binding (i.e., exhibits at least or at least about 10 7 -10 8 "Functional fragments" or "analogs of anti-MASP-2 antibodies" are fragments or analogs that can prevent or substantially reduce the ability of the receptor to bind to a ligand or initiate signal transduction. As used herein, functional fragments are generally synonymous with "antibody fragments" and, with respect to antibodies, can refer to fragments that can prevent or substantially reduce the ability of the receptor to bind to a ligand or initiate signal transduction, such as Fv, Fab, F(ab') 2 The "Fv" fragment is a dimer formed by the variable domain of a heavy chain and the variable domain of a light chain by non-covalent bonding (V H -V L In this configuration, the three CDRs of each variable domain interact to determine the V H -V L The target binding site on the surface of the dimer is the same as that of the complete antibody. The six CDRs together confer the target binding specificity of the complete antibody. However, even a single variable domain (or half of an Fv comprising only three target-specific CDRs) can still have the ability to recognize and bind to the target.

[0038] As used herein, the term "bispecific" (Bispecific antibody, BsAb) refers to antibodies and / or antigen binding molecules that can specifically bind to two different antigenic determinants. Typically, bispecific antibodies and / or antigen binding molecules contain two antigen binding sites, each of which is specific for different antigenic determinants. In certain embodiments, the bispecific antibody and / or antigen binding molecule can simultaneously bind to two antigenic determinants, particularly two antigenic determinants expressed on two different cells.

[0039] As used herein, "monoclonal antibody" refers to a population of identical antibodies, meaning that each individual antibody molecule in a monoclonal antibody population is identical to other antibody molecules. This property is in contrast to the properties of a polyclonal population of antibodies, which contains antibodies with a variety of different sequences. Monoclonal antibodies can be prepared by many well-known methods (Smith et al. (2004) J. Clin. Pathol. 57, 912-917; and Nelson et al., J Clin Pathol (2000), 53, 111-117). For example, monoclonal antibodies can be prepared by immortalized B cells, for example, by fusion with myeloma cells to produce hybridoma cell lines or by infecting B cells with viruses such as EBV. Recombinant technology can also be used to prepare antibodies from a clonal population of host cells in vitro by transforming host cells with a plasmid carrying an artificial sequence of nucleotides encoding the antibody.

[0040] As used herein, the term "hybridoma" or "hybridoma cell" refers to a cell or cell line (usually a myeloma or lymphoma cell) produced by the fusion of an antibody-producing lymphocyte and a non-antibody-producing cancer cell. As known to those of ordinary skill in the art, hybridomas can proliferate and continue to produce a specific monoclonal antibody. Methods for producing hybridomas are known in the art (see, e.g., Harlow & Lane, 1988). When referring to the term "hybridoma" or "hybridoma cell", it also includes subclones and progeny cells of the hybridoma.

[0041] As used herein, a full-length antibody is an antibody having two full-length heavy chains (e.g., VH-CH1-CH2-CH3 or VH-CH1-CH2-CH3-CH4) and two full-length light chains (VL-CL) and a hinge region, such as antibodies naturally produced by antibody-secreting B cells and antibodies produced synthetically with the same domains.

[0042] The term "chimeric antibody" refers to antibodies in which the variable region sequences are derived from one species and the constant region sequences are derived from another species, such as antibodies in which the variable region sequences are derived from mouse antibodies and the constant region sequences are derived from human antibodies.

[0043] "Humanized" antibodies refer to non-human (eg, mouse) antibody forms that are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (eg, Fv, Fab, Fab', F(ab') 2 Humanized antibodies are human immunoglobulins (recipient antibodies) in which residues in the complementary determining regions (CDRs) of the recipient antibody are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity.

[0044] In addition, in humanization, it is also possible to mutate the amino acid residues in the CDR1, CDR2 and / or CDR3 regions of VH and / or VL, thereby improving one or more binding properties (e.g., affinity) of the antibody. For example, PCR-mediated mutations can be used to introduce mutations, and their effects on antibody binding or other functional properties can be evaluated using in vitro or in vivo tests described herein. Typically, conservative mutations are introduced. Such mutations can be amino acid substitutions, additions or deletions. In addition, the mutations in the CDR are generally no more than one or two. Therefore, the humanized antibodies described in the present disclosure also encompass antibodies comprising 1 or 2 amino acid mutations in the CDR.

[0045] As used herein, the term "CDR" refers to a complementarity-determining region, and each heavy chain and light chain of a known antibody molecule has three CDRs. CDRs are also referred to as hypervariable regions and are present in the variable regions of each heavy chain and light chain of an antibody, with very high variability sites in the primary structure of the CDRs. In this specification, the CDRs of the heavy chain are represented by CDR1, CDR2, and CDR3 from the amino terminal of the amino terminal sequence of the heavy chain, and the CDRs of the light chain are represented by CDR1, CDR2, and CDR3 from the amino terminal of the amino terminal sequence of the light chain. These sites are adjacent to each other in the tertiary structure and determine the specificity of the antigen to which the antibody binds.

[0046] As used herein, the term "epitope" refers to any antigenic determinant on an antigen to which the paratope of an antibody binds. Epitopic determinants usually comprise chemically active surface patterns of molecules, such as amino acids or sugar side chains, and usually have specific three-dimensional structural characteristics as well as specific charge characteristics.

[0047] As used herein, "specific binding" or "immunospecifically binding" with respect to an antibody or antigen-binding fragment thereof are used interchangeably herein and refer to the ability of an antibody or antigen-binding fragment to form one or more non-covalent bonds with a cognate antigen through non-covalent interactions between the antibody combining sites of the antibody and the antigen. The antigen may be an isolated antigen or present in a tumor cell. Typically, an antibody that immunospecifically binds (or specifically binds) to an antigen is present in an amount of about or 1×10 7 M -1 or 1x10 8 M -1 or greater affinity constant Ka (or 1x10 -7 M or 1×10 -8M or lower dissociation constant (Kd)) binds to the antigen. Affinity constants can be determined by standard kinetic methods of antibody reactions, e.g., immunoassays, surface plasmon resonance (SPR) (Rich and Myszka (2000) Curr. Opin. Biotechnol 11:54; Englebienne (1998) Analyst. 123:1599), isothermal titration calorimetry (ITC), or other kinetic interaction assays known in the art (see, e.g., Paul, ed., Fundamental Immunology, 2nd ed., Raven Press, New York, pages 332-336 (1989); also see U.S. Pat. No. 7,229,619 describing exemplary SPR and ITC methods for calculating the binding affinity of antibodies). Instruments and methods for real-time detection and monitoring of binding rates are known and commercially available (see, BiaCore 2000, Biacore AB, Upsala, Sweden and GE Healthcare Life Sciences; Malmqvist (2000) Biochem. Soc. Trans. 27:335).

[0048] As used herein, the terms "polynucleotide" and "nucleic acid molecule" refer to an oligomer or polymer comprising at least two linked nucleotides or nucleotide derivatives, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), usually linked together by a phosphodiester bond. As used herein, the term "nucleic acid molecule" is intended to include DNA molecules and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded, and can be cDNA.

[0049] As used herein, an isolated nucleic acid molecule is a nucleic acid molecule isolated from other nucleic acid molecules present in the natural source of the nucleic acid molecule. "Isolated" nucleic acid molecules such as cDNA molecules can be substantially free of other cellular materials or culture media when prepared by recombinant technology, or substantially free of chemical precursors or other chemical compositions when chemosynthesized. The exemplary isolated nucleic acid molecules provided herein include the isolated nucleic acid molecules of the antibody or Fab provided by encoding.

[0050] As used herein, "operably linked" with respect to a nucleic acid sequence, region, element or domain means that the nucleic acid regions are functionally related to each other. For example, a promoter can be operably linked to a nucleic acid encoding a polypeptide so that the promoter regulates or mediates transcription of the nucleic acid.

[0051] Also provided are "conservative sequence modifications" of the sequences described in the sequence listings described herein, i.e., nucleotide and amino acid sequence modifications that do not eliminate the binding of the antibody encoded by the nucleotide sequence or containing the amino acid sequence to the antigen. These conservative sequence modifications include conservative nucleotide and amino acid substitutions and nucleotide and amino acid additions and deletions. For example, modifications can be introduced into the sequence listings described herein by standard techniques known in the art (e.g., site-directed mutagenesis and PCR-mediated mutagenesis). Conservative sequence modifications include conservative amino acid substitutions, in which amino acid residues are replaced with amino acid residues with similar side chains. Families of amino acid residues with similar side chains are already defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), amino acids with beta-branched side chains (e.g., threonine, valine, isoleucine), and amino acids with aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted nonessential amino acid residue in an anti-MASP-2 antibody is preferably replaced with another amino acid residue from the same side chain family. Methods for identifying conservative nucleotide and amino acid substitutions that do not eliminate antigen binding are well known in the art (e.g., see Brummell et al., Biochem. 32: 1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10): 879-884 (1999); Burks et al., Proc. Natl. Acad. Sci. USA 94: 412-417 (1997)).

[0052] Alternatively, in another embodiment, mutations can be randomly introduced along all or a portion of an anti-MASP-2 antibody coding sequence by, for example, saturation mutagenesis, and the resultant modified anti-MASP-2 antibodies can be screened for improved binding activity.

[0053] As used herein, "expression" refers to the process of producing a polypeptide by transcription and translation of a polynucleotide. The expression level of a polypeptide can be evaluated using any method known in the art, including, for example, methods for determining the amount of polypeptide produced from a host cell. Such methods may include, but are not limited to, quantifying polypeptides in cell lysates by ELISA, Coomassie blue staining after gel electrophoresis, Lowry protein assay, and Bradford protein assay.

[0054] As used herein, "host cell" is a cell that is used to receive, maintain, replicate and amplify a vector. Host cells can also be used to express polypeptides encoded by the vector. When the host cell divides, the nucleic acid contained in the vector replicates, thereby amplifying the nucleic acid. The host cell can be a eukaryotic cell or a prokaryotic cell. Suitable host cells include, but are not limited to, CHO cells, various COS cells, HeLa cells, HEK cells such as HEK 293 cells.

[0055] As used herein, "vector" is a replicable nucleic acid from which one or more heterologous proteins can be expressed when the vector is transformed into an appropriate host cell. Vectors include those into which nucleic acids encoding polypeptides or fragments thereof can be introduced, usually by restriction digestion and ligation. Vectors also include those containing nucleic acids encoding polypeptides. Vectors are used to introduce nucleic acids encoding polypeptides into host cells, for amplification of nucleic acids or for expression / display of polypeptides encoded by nucleic acids. Vectors are usually kept free, but can be designed to integrate genes or parts thereof into chromosomes of the genome. Artificial chromosome vectors are also contemplated, such as yeast artificial vectors and mammalian artificial chromosomes. The selection and use of such vehicles are well known to those skilled in the art.

[0056] As used herein, vectors also include “viral vectors” or “viral vectors.” Viral vectors are engineered viruses that are operably linked to exogenous genes to transfer (as a vehicle or shuttle) the exogenous genes into cells.

[0057] As used herein, "expression vector" includes vectors capable of expressing DNA, which is operably linked to regulatory sequences such as promoter regions that can affect the expression of such DNA fragments. Such additional fragments may include promoter and terminator sequences, and may optionally include one or more replication origins, one or more selection markers, enhancers, polyadenylation signals, etc. Expression vectors are generally derived from plasmid or viral DNA, or may contain elements of both. Therefore, expression vectors refer to recombinant DNA or RNA constructs, such as plasmids, phages, recombinant viruses or other vectors, which, when introduced into appropriate host cells, result in the expression of cloned DNA. Suitable expression vectors are well known to those skilled in the art, and include expression vectors that are replicable in eukaryotic cells and / or prokaryotic cells and expression vectors that remain free or are integrated into the host cell genome.

[0058] As used herein, "treating" an individual suffering from a disease or condition means that the symptoms of the individual are partially or completely relieved, or remain unchanged after treatment. Therefore, treatment includes prevention, treatment and / or cure. Prevention refers to preventing potential diseases and / or preventing symptoms from worsening or disease development. Treatment also includes any pharmaceutical use of any antibody or antigen-binding fragment thereof provided and any composition provided herein.

[0059] As used herein, "therapeutic effect" refers to the effect resulting from treatment of a subject that alters, typically ameliorates or improves the symptoms of a disease or condition, or cures the disease or condition.

[0060] As used herein, "therapeutically effective amount" or "therapeutically effective dose" refers to an amount of a substance, compound, material, or composition comprising a compound that is at least sufficient to produce a therapeutic effect after administration to a subject. Therefore, it is the amount necessary to prevent, cure, improve, block, or partially block the symptoms of a disease or disorder.

[0061] As used herein, a "prophylactically effective amount" or "prophylactically effective dose" refers to an amount of a substance, compound, material, or composition comprising a compound that, when administered to a subject, will have the desired prophylactic effect, e.g., preventing or delaying the onset or recurrence of a disease or symptom, reducing the likelihood of the onset or recurrence of a disease or symptom. A complete prophylactically effective dose need not occur by administering one dose, and may occur only after administering a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations.

[0062] As used herein, the term "patient" or "subject" refers to a mammal, such as a human.

[0063] II. Detailed description of specific implementation scheme

[0064] In one aspect, the present disclosure provides an antibody or antigen binding portion thereof that binds to MASP-2 and comprises a heavy chain CDR selected from the group consisting of the amino acid sequences of SEQ ID NO. 7, 8, 9, 17, 18, 26, 27, 28, 36, 37, 38, 46, 47, 60, 65, 66, 67, 79, 85, 86, 94, 95, 96, 104, 105, 113, 114, 126, 127, 128, 136, 137, 138, 141, 149, 150, 151, 165, 183, 192, or any variant thereof, and / or a heavy chain CDR selected from the group consisting of the amino acid sequences of SEQ ID NO. NO. 12, 13, 14, 21, 22, 23, 31, 32, 33, 41, 42, 43, 50, 51, 52, 57, 70, 71, 72, 82, 91, 108, 109, 110, 117, 118, 123, 131, 132, 133, 144, 145, 146, 154, 155, 156, 161, 162, 168, 173, 174, 201, 202, 207, 216 or any variant thereof.

[0065] The antibody or antigen-binding portion thereof according to the preceding aspect comprises a heavy chain CDR1 selected from the group consisting of the amino acid sequence of SEQ ID NO.7, 26, 36, 65, 94, 126, 136, 149 or any variant thereof, a heavy chain CDR2 selected from the group consisting of the amino acid sequence of SEQ ID NO.8, 17, 27, 37, 46, 66, 85, 95, 104, 113, 127, 137, 150, 165, 183, 192 or any variant thereof, a heavy chain CDR3 selected from the group consisting of the amino acid sequence of SEQ ID NO.9, 18, 28, 38, 47, 60, 67, 79, 86, 96, 105, 114, 128, 138, 141, 151 or any variant thereof; and / or a heavy chain CDR3 selected from the group consisting of the amino acid sequence of SEQ ID NO. NO.12, 21, 31, 41, 50, 70, 99, 108, 117, 123, 131, 144, 154, 161, 201, 216 or any variant thereof, a light chain CDR1 selected from the amino acid sequence SEQ ID NO.13, 22, 32, 42, 51, 57, 71, 100, 109, 118, 132, 145, 155, 162, 168, 173, 202, 207 or any variant thereof, a light chain CDR3 selected from the amino acid sequence SEQ ID NO.14, 23, 33, 43, 52, 72, 82, 91, 101, 110, 133, 146, 156, 174 or any variant thereof.

[0066] The antibody or antigen binding portion thereof according to the previous aspect comprises a combination of heavy and light chain CDRs selected from:

[0067] (1) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 8, 9, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 12, 13, 14, respectively;

[0068] (2) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 18, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 21, 22, 23, respectively;

[0069] (3) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 26, 27, 28, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 31, 32, 33, respectively;

[0070] (4) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 36, 37, 38, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 41, 42, 43, respectively;

[0071] (5) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 46, 47, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 50, 51, 52, respectively;

[0072] (6) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 18, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 21, 57, 23, respectively;

[0073] (7) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 60, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 50, 51, 52, respectively;

[0074] (8) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 65, 66, 67, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 70, 71, 72, respectively;

[0075] (9) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 47, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 50, 51, 52, respectively;

[0076] (10) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 79, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 50, 51, 82, respectively;

[0077] (11) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 36, 85, 86, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 41, 42, 43, respectively;

[0078] (12) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 36, 85, 86, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 41, 42, 91, respectively;

[0079] (13) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 94, 95, 96, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 99, 100, 101, respectively;

[0080] (14) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 104 and 105, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 108, 109 and 110, respectively;

[0081] (15) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 26, 113, 114, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 117, 118, 33, respectively;

[0082] (16) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 94, 95, 96, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 123, 100, 101, respectively;

[0083] (17) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 126, 127, 128, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 131, 132, 133, respectively;

[0084] (18) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 136, 137, 138, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 131, 132, 133, respectively;

[0085] (19) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 141, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 144, 145, 146, respectively;

[0086] (20) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 149, 150, 151, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 154, 155, 156, respectively;

[0087] (21) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 9, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 161, 162, 14, respectively;

[0088] (22) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 165 and 9, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 144, 168 and 14, respectively;

[0089] (23) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 9, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 144, 173, 174, respectively;

[0090] (24) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 136, 183, 138, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 216, 132, 133, respectively;

[0091] (25) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 126, 192, 128, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 216, 132, 133, respectively;

[0092] (26) heavy chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 7, 17, 141, respectively, and / or light chain CDR1, CDR2 and CDR3 sequences comprising SEQ ID NOs: 201, 202, 146, respectively;

[0093] (27) The heavy chain CDR1, CDR2 and CDR3 sequences respectively comprise SEQ ID NOs: 7, 17, 141, and / or the light chain CDR1, CDR2 and CDR3 sequences respectively comprise SEQ ID NOs: 201, 207, 146.

[0094] The antibody or antigen-binding portion thereof according to the preceding aspect comprises a heavy chain variable region selected from the group consisting of the amino acid sequence of SEQ ID NO. 5, 15, 24, 34, 44, 53, 58, 63, 73, 77, 83, 92, 102, 111, 119, 124, 134, 139, 147, 157, 163, 169, 175, 177, 179, 181, 184, 186, 188, 190, 193, 195, 197 or any variant thereof, and / or a heavy chain variable region selected from the group consisting of the amino acid sequence of SEQ ID NO. NO.10, 19, 29, 39, 48, 55, 61, 68, 75, 80, 87, 89, 97, 106, 115, 121, 129, 142, 152, 159, 166, 171, 199, 203, 205, 208, 210, 212, 214 or any variant thereof;

[0095] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.5 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.10 or any variant thereof;

[0096] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.15 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.19 or any variant thereof;

[0097] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.24 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.29 or any variant thereof;

[0098] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.34 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.39 or any variant thereof;

[0099] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.44 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.48 or any variant thereof;

[0100] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.53 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.55 or any variant thereof;

[0101] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.58 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.61 or any variant thereof;

[0102] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.63 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.68 or any variant thereof;

[0103] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.73 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.75 or any variant thereof;

[0104] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.77 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.80 or any variant thereof;

[0105] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.83 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.87 or any variant thereof;

[0106] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.83 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.89 or any variant thereof;

[0107] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.92 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.97 or any variant thereof;

[0108] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.102 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.106 or any variant thereof;

[0109] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.111 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.115 or any variant thereof;

[0110] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.119 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.121 or any variant thereof;

[0111] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.124 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.129 or any variant thereof;

[0112] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.134 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.129 or any variant thereof;

[0113] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.139 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.142 or any variant thereof;

[0114] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.147 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.152 or any variant thereof;

[0115] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.157 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.159 or any variant thereof;

[0116] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.163 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.166 or any variant thereof;

[0117] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.169 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.171 or any variant thereof;

[0118] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.181 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0119] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.184 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0120] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.186 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0121] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.188 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0122] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.190 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0123] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.193 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0124] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.195 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0125] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.197 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.214 or any variant thereof;

[0126] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.175 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.199 or any variant thereof;

[0127] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.175 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.203 or any variant thereof;

[0128] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.175 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.205 or any variant thereof;

[0129] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.177 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.205 or any variant thereof;

[0130] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.177 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.199 or any variant thereof;

[0131] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.177 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.203 or any variant thereof;

[0132] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.179 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.199 or any variant thereof;

[0133] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.179 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.203 or any variant thereof;

[0134] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.179 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.205 or any variant thereof;

[0135] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.175 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.208 or any variant thereof;

[0136] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.175 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.210 or any variant thereof;

[0137] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.175 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.212 or any variant thereof;

[0138] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.177 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.208 or any variant thereof;

[0139] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.177 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.210 or any variant thereof;

[0140] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.177 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.212 or any variant thereof;

[0141] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.179 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.208 or any variant thereof;

[0142] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region of the amino acid sequence SEQ ID NO.179 or any variant thereof, and a light chain variable region of the amino acid sequence SEQ ID NO.210 or any variant thereof;

[0143] In some preferred embodiments, the antibody or antigen-binding portion thereof comprises a heavy chain variable region having an amino acid sequence of SEQ ID NO.179 or any variant thereof, and a light chain variable region having an amino acid sequence of SEQ ID NO.212 or any variant thereof.

[0144] In one aspect, the present disclosure provides a bispecific or multispecific molecule comprising the antibody or antigen-binding portion thereof of any of the preceding aspects.

[0145] In some preferred embodiments of the present disclosure, the antibody or antigen-binding portion thereof is humanized.

[0146] In one aspect, the disclosure provides a nucleic acid molecule encoding an antibody or antigen binding portion thereof, or a bispecific or multispecific molecule according to any of the preceding aspects.

[0147] In some preferred embodiments, the nucleic acid comprises an antibody heavy chain variable region nucleic acid sequence selected from SEQ ID NO. 6, 16, 25, 35, 45, 54, 59, 64, 74, 78, 84, 93, 103, 112, 120, 125, 135, 140, 148, 158, 164, 170, 176, 178, 180, 182, 185, 187, 189, 191, 194, 196, 198 or any variant thereof, and / or an antibody heavy chain variable region nucleic acid sequence selected from SEQ ID NO. ID NO.11, 20, 30, 40, 49, 56, 62, 69, 76, 81, 88, 90, 98, 107, 116, 122, 130, 143, 153, 160, 167, 172, 200, 204, 206, 209, 211, 213, 215 or any variant thereof;

[0148] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 6 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 11 or any variant thereof;

[0149] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 16 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 20 or any variant thereof;

[0150] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 25 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 30 or any variant thereof;

[0151] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 35 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 40 or any variant thereof;

[0152] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 45 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 49 or any variant thereof;

[0153] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 54 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 56 or any variant thereof;

[0154] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 59 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 62 or any variant thereof;

[0155] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 64 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 69 or any variant thereof;

[0156] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 74 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 76 or any variant thereof;

[0157] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 78 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 81 or any variant thereof;

[0158] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 84 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 88 or any variant thereof;

[0159] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 84 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 90 or any variant thereof;

[0160] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 93 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 98 or any variant thereof;

[0161] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 103 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 107 or any variant thereof;

[0162] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 112 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 116 or any variant thereof;

[0163] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 120 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 122 or any variant thereof;

[0164] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 125 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 130 or any variant thereof;

[0165] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 135 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 130 or any variant thereof;

[0166] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 140 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 143 or any variant thereof;

[0167] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 148 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 153 or any variant thereof;

[0168] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 158 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 160 or any variant thereof;

[0169] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 164 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 167 or any variant thereof;

[0170] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 170 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 172 or any variant thereof;

[0171] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 182 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0172] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 185 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0173] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 187 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0174] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 189 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0175] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 191 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0176] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 194 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0177] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 196 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0178] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 198 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 215 or any variant thereof;

[0179] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 176 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 200 or any variant thereof;

[0180] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 176 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 204 or any variant thereof;

[0181] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 176 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 206 or any variant thereof;

[0182] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 178 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 206 or any variant thereof;

[0183] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 178 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 200 or any variant thereof;

[0184] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 178 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 204 or any variant thereof;

[0185] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 180 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 200 or any variant thereof;

[0186] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 180 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 204 or any variant thereof;

[0187] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 180 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 206 or any variant thereof;

[0188] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 176 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 209 or any variant thereof;

[0189] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 176 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 211 or any variant thereof;

[0190] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 176 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 213 or any variant thereof;

[0191] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 178 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 209 or any variant thereof;

[0192] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 178 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 211 or any variant thereof;

[0193] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 178 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 213 or any variant thereof;

[0194] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 180 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 209 or any variant thereof;

[0195] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 180 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 211 or any variant thereof;

[0196] In some preferred embodiments, the nucleic acid comprises a heavy chain variable region nucleotide sequence of SEQ ID NO: 180 or any variant thereof, and a light chain variable region nucleotide sequence of SEQ ID NO: 213 or any variant thereof.

[0197] In one aspect, the disclosure provides antibodies, or antigen binding portions thereof, that bind to MASP-2 and have at least greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity to the antibody, or antigen binding portion thereof, of any of the preceding aspects.

[0198] In one aspect, the present disclosure provides a nucleic acid molecule encoding an antibody or antigen-binding portion thereof as described in any of the preceding aspects, or a nucleic acid molecule having at least greater than 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity thereto.

[0199] In one aspect, the present disclosure provides a vector comprising a nucleic acid as in any of the preceding aspects.

[0200] In one aspect, the present disclosure provides a cell comprising a vector as in any of the preceding aspects.

[0201] In one aspect, the present invention discloses a composition comprising the aforementioned antibody or antigen-binding portion thereof, bispecific or multispecific molecule, nucleic acid, rights vector and / or cell.

[0202] The antibodies of the present disclosure are useful as therapeutic or diagnostic tools in a variety of diseases in which MASP-2 is adversely expressed or found.

[0203] The method of treating a disease associated with MASP-2-dependent complement activation with the MASP-2 inhibitory agent of the present disclosure comprises the step of administering to the subject a therapeutically effective amount of the antibody or antigen-binding fragment thereof or nucleic acid molecule or vector or cell or pharmaceutical composition of any of the aforementioned aspects.

[0204] MASP-2-dependent complement activation has been implicated as contributing to the pathogenesis of a number of acute and chronic disease states, including MASP-2-dependent complement-mediated vascular conditions, ischemia-reperfusion injury, atherosclerosis, inflammatory gastrointestinal disorders, pulmonary conditions, extracorporeal reperfusion procedures, skeletal muscle conditions, renal conditions, skin conditions, organ or tissue transplantation, nervous system disorders or injuries, blood disorders, genitourinary tract conditions, diabetes, chemotherapy or radiation treatment, malignancies, endocrine disorders, coagulation disorders, or ophthalmic conditions. Thus, the MASP-2 inhibitory antibodies of the present disclosure may be used to treat the above-mentioned diseases and conditions.

[0205] The clinical condition of a preferred embodiment of the present disclosure is a chronic inflammatory disease. The chronic inflammatory disease may be, for example, an autoimmune inflammatory condition.

[0206] Autoimmune inflammatory conditions (also referred to herein as "autoimmune disorders") can be loosely categorized into those that are primarily localized to a particular organ or tissue and those that affect the entire body. Examples of organ-specific disorders (and affected organs) include multiple sclerosis (myelin coating on nerve processes), type I diabetes (pancreas), Hashimoto's thyroiditis (thyroid), pernicious anemia (stomach), Addison's disease (adrenal glands), myasthenia gravis (acetylcholine receptors on neuromuscular junctions), rheumatoid arthritis (joint linings), uveitis (eyes), psoriasis (skin), Guillain-Barre syndrome (nerve cells), and Graves' disease (thyroid). Systemic autoimmune diseases include systemic lupus erythematosus, glomeronephritis, and dermatomyositis.

[0207] In one embodiment of the present disclosure, the preferred clinical condition is selected from rheumatoid arthritis or systemic lupus erythematosus.

[0208] Other examples of autoimmune disorders include asthma, eczema, atopic dermatitis, contact dermatitis, other eczematous dermatitis, seborrheic dermatitis, rhinitis, lichen planus, pemplugus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitis, erythema, eosinophilia, alopecia areata, arteriosclerosis, primary biliary cirrhosis, and nephrotic syndrome. Related diseases include enteritis, such as coeliac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, inflammatory bowel disease, Chrohn's disease, and ulcerative colitis, as well as food-related allergies.

[0209] In another embodiment of the present disclosure, the clinical condition is characterized by massive cell loss, for example due to programmed cell death or necrosis. The necrosis or programmed cell death can be induced by a variety of factors.

[0210] In a preferred embodiment of the present disclosure, the clinical condition is an ischemic / reperfusion injury. Ischemia can be caused by various reasons, such as after a stroke, myocardial infarction, major surgery or organ transplantation. Thus, the clinical condition can be an ischemic / reperfusion injury caused by, for example, a stroke, myocardial infarction, major surgery or organ transplantation.

[0211] Thus, the clinical condition may be an ischemic / reperfusion injury that is the result of PTCA (percutaneous transluminal coronary angioplasty) or CABG (coronary artery bypass grafting). In addition, the clinical condition may be an ischemic / reperfusion injury that is the result of acute myocardial infarction or cerebral ischemia.

[0212] The dosage of the medicament comprising the MASP-2 inhibitory agent generally varies depending on the age, weight, height, sex, general disease condition and medical history of the subject. For example, the MASP-2 inhibitory agent, such as a MASP-2 antibody, can be administered in a dosage range of about 0.010-10.0 mg / kg, preferably 0.010-1.0 mg / kg, more preferably 0.010-0.1 mg / kg of the subject's body weight. In certain embodiments, the composition comprises a combination of an anti-MASP-2 antibody and a MASP-2 inhibitory peptide.

[0213] Compositions and methods comprising a MASP-2 inhibitory agent may optionally include one or more additional therapeutic agents that may enhance the activity of the MASP-2 inhibitory agent or provide related therapeutic functions in an additive or synergistic manner. For example, one or more MASP-2 inhibitory agents may be used in combination with one or more anti-inflammatory and / or analgesic agents. The amount and selection of additional agents may be determined to achieve the desired therapeutic result. Suitable anti-inflammatory and / or analgesic drugs include: 5-hydroxytryptamine receptor antagonists; 5-hydroxytryptamine receptor agonists; histamine receptor antagonists; bradykinin receptor antagonists; kallikrein inhibitors; tachykinin receptor antagonists, including neurokinin 1 and neurokinin 2 receptor subtype antagonists; calcitonin gene-related peptide (CGRP) receptor antagonists; interleukin receptor antagonists; inhibitors of enzymes active in the arachidonic acid metabolite synthesis pathway, including phospholipase inhibitors, including PLA2 isoform inhibitors and PLCγ isoform inhibitors, cyclooxygenase (COX) inhibitors (which may be COX-1, COX-2 inhibitors or non-selective COX-1 and COX-2 inhibitors), lipoxygenase inhibitors; 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; purine receptor agonists and antagonists, including P2x receptor antagonists and P2Y receptor agonists; adenosine triphosphate (ATP)-sensitive potassium channel openers; MAP kinase inhibitors; nicotinic acid acetylcholine inhibitors; and alpha adrenergic receptor agonists (including alpha-1, alpha-2 and non-selective alpha-1 and alpha-2 agonists).

[0214] When used to inhibit or treat restenosis, the MASP-2 inhibitory agents of the present disclosure may be co-administered in combination with one or more anti-restenotic drugs. Suitable anti-restenotic drugs include: antiplatelet agents, including thrombin inhibitors and receptor antagonists, adenosine diphosphate (ADP) receptor antagonists (also known as purinergic receptor 1 receptor antagonists), thromboxane inhibitors and receptor antagonists, and platelet membrane glycoprotein receptor antagonists; inhibitors of cell adhesion molecules, including selectin inhibitors and integrin free radicals; anti-chemotactic agents; interleukin receptor antagonists; and intracellular signal transduction inhibitors, including protein kinase C (PKC) inhibitors and protein tyrosine phosphatases, modulators of intracellular protein tyrosine kinase inhibitors, inhibitors of src homology 2 (SH2) domains, and calcium channel antagonists.

[0215] The MASP-2 inhibitory agents of the present disclosure may also be used in combination with one or more other complement inhibitors. Currently no complement inhibitors are approved for use in humans, however some drugs have been shown to block complement in vivo. Many of these drugs are also toxic, or are only partial inhibitors, and the use of these drugs is limited to research tools. K76COOH and nafamostat mesylate are two drugs that have some efficacy in transplant animal models. Studies have shown that low molecular weight heparins are also effective in modulating complement activity. We believe that these small molecule inhibitors may be used as drugs in combination with the MASP-2 inhibitors of the present disclosure.

[0216] Other naturally occurring complement inhibitors can be used in combination with the MASP-2 inhibitory agents of the present disclosure. Biological inhibitors of complement include soluble complement factor 1 (sCR1). This is a naturally occurring inhibitor found on the outer membrane of human cells. Other membrane inhibitors include DAF, MCP, and CD59. Recombinant forms have been assayed for anti-complement activity in vitro and in vivo. sCR1 has been shown to be effective in xenotransplantation, where the complement system (both alternative and classical) causes an overreactive rejection syndrome within minutes of perfusing the newly transplanted organ with blood. Use of sCR1 protects and prolongs the survival of the transplanted organ, indicating a complement pathway in the mechanism of organ survival.

[0217] Other complement inhibitors suitable for use in combination with the compositions of the present disclosure include, for example, monoclonal antibodies (MoAbs) being developed by Alexion Pharmaceuticals, Inc. New Haven, Connecticut, and anti-properdin monoclonal antibodies (MoAbs).

[0218] When used to treat arthritis (e.g., osteoarthritis and rheumatoid arthritis), the MASP-2 inhibitory agents of the present disclosure may be used in combination with one or more chondroprotective agents, which may include one or more promoters of cartilage anabolism and / or one or more inhibitors of cartilage catabolism, including, where appropriate, both anabolicagents and catabolic inhibitory agents administered simultaneously. Suitable anabolic chondroprotective agents include interleukin (IL) receptor agonists, including IL-4, IL-10, IL-13, rhIL-4, rhIL-10 and rhIL-13, and chimeric IL-4, IL-10 or IL-13; transforming growth factor β superfamily agonists (including TGF-β, TGF-β1, TGF-β2, TGF-β3), bone morphogenic proteins including BMP-2, BMP-4, BMP-5, BMP-6, BMP-7 (OP-1) and OP-2 / BMP-8, growth differentiation factors including GDF-5, GDF-6 and GDF-7, recombinant TGF-β and BMP, and chimeric TGF-β and BMP; insulin-like growth factors, including IGF-1; and fibroblast growth factors, including bFGF. Suitable chondroprotective agents that inhibit catabolism include interleukin-1 (IL-1) receptor antagonists (IL-1ra), including soluble human IL-1 receptor (shuIL-1R), rshuIL-1R, rhIL-1ra, anti-IL1 antibodies, AF11567 and AF12198; tumor necrosis factor (TNF) receptor antagonists (TNF-α), including soluble receptors including sTNFR1 and sTNFRII, recombinant TNF soluble receptors and chimeric TNF soluble receptors, including chimeric rhTNFR:Fc, Fc fusion soluble receptors and anti-TNF antibodies; cyclooxygenase- 2 (COX-2 specific) inhibitors; including DuP697, SC-58451, celecoxib, rofecoxib, nimesulide, diclofenac, meloxicam, piroxicam, NS-398, RS-57067, SC-57666, SC-58125, flosulide, etodolac, L-745,337, and DFU-T-614;Mitogen-activated protein kinase (MAPK) inhibitors, including inhibitors of ERKi, ERK2, SAPK1, SAPK2a, SAPK2b, SAPK2d, SAPK3, including SB 203580, SB 203580 iodine, SB202190, SB 242235, SB 220025, RWJ 67657, RWJ68354, FR 133605, L-167307, PD 98059, PD 1693 16; nuclear factor κB (NFκB) inhibitors, including caffeic acid phenethyl ester (CAPE), DM-CAPE, SN-50 peptide, hymenialdisine and pyrrolidone dithiocarbamate; nitric oxide synthase (NOS) inhibitors, including NG-monomethyl-L-arginine, 1400W, diphenylene iodide, S-methylisothiourea, S-(aminoethyl)isothiourea, L-N6-(1-iminoethyl)lysine, 1,3-PBITU, 2-ethyl-2-thiopseudurea, aminoguanidine, Nω-nitro-L-arginine, Nω-nitro-L-arginine methyl ester; inhibitors of matrix metalloproteinases (MMPs), including MMP-1, MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-1 1. Inhibitors of MMP-12, MMP-13, MMP-14 and MMP-15, including U-24522, minocycline, 4-Abz-Gly-Pro-D-Leu-D-Ala-NHOH, Ac-Arg-Cys-Gly-Val-Pro-Asp-NH2, recombinant human TIMP1, recombinant human TIMP2 and phosphoramidon; cell adhesion molecules, including integrin agonists and antagonists, including αVβ3 MoAb LM 609 and echistatin; anti-chemotactic agents, including F-Met-Leu-Phe receptor, IL-8 receptor, MCP-1 receptor and MIP1-I / RANTES receptor; intracellular signal transduction inhibitors, including (a) protein kinase inhibitors, including (i) protein kinase C (PKC) inhibitors (isoenzymes) including calphostin C C), G-6203 and GF109203X and (ii) protein tyrosine kinase inhibitors; (b) intracellular protein tyrosine phosphatase (PTPase) regulators; and (c) SH2 domain inhibitors (src homology 2 domain). ;

[0219] For some applications, it may be advantageous to combine a MASP-2 inhibitory agent of the present disclosure with a spasm inhibitor. For example, for genitourinary applications, it may be advantageous to include at least one smooth muscle spasm inhibitor and / or at least one anti-inflammatory agent, and for vascular surgery, it may be beneficial to include at least one vasospasm inhibitor and / or at least one anti-inflammatory agent and / or at least one anti-restenotic agent. Suitable examples of spasm inhibitors include: 5-hydroxytryptamine 2 receptor subtype antagonists; tachykinin receptor antagonists; nitric oxide donors; ATP-sensitive potassium channel openers; calcium channel antagonists; and endothelin receptor antagonists.

[0220] In one embodiment of the present disclosure, the antibody is administered in a single dose or in multiple doses.

[0221] If the antibody is administered in multiple doses according to the present disclosure, it is preferably administered in at least 3 doses, at least 4 doses, at least 5 doses, at least 6 doses, at least 7 doses, at least 8 doses, at least 9 doses or at least 10 doses and preferably up to 30 doses, 25 doses, 20 doses, 15 doses or 10 doses. The doses of the antibody are preferably administered at intervals of at least 7 days, at least 10 days, at least 14 days or at least 20 days. The doses of the antibody are preferably administered at intervals of 7 to 30 days, 10 to 20 days and preferably about 14 days.

[0222] In one aspect, the disclosure provides use of an antibody or antigen-binding fragment thereof or a nucleic acid molecule or a vector or a cell or a pharmaceutical composition of any of the preceding aspects for the preparation of a medicament for treating a disease associated with MASP-2-dependent complement activation in a subject.

[0223] In one aspect, the present disclosure provides a kit comprising the aforementioned antibody or antigen-binding portion thereof, bispecific or multispecific molecule, nucleic acid molecule, vector, cell and / or composition.

[0224] The kit disclosed herein includes antibodies, fragments thereof, homologues thereof, derivatives thereof, etc. disclosed herein, such as conjugates with labels or cytotoxicity, and instructions for use of antibodies, conjugates that kill specific types of cells, etc. The instructions may include instructions for using antibodies, conjugates, etc. in vitro, in vivo, or ex vivo. The antibody may be in liquid form or solid, usually lyophilized. The kit may contain other suitable reagents, such as buffers, reconstitution solutions, and other necessary components for the intended use. It is contemplated that the reagent combination packaged in a predetermined amount is combined with instructions for its use, such as for therapeutic use or for performing diagnostic assays. When the antibody is labeled, such as with an enzyme label, the kit may include substrates and cofactors required by the enzyme (e.g., substrate precursors that provide detectable chromophores or fluorophores). In addition, other additives, such as stabilizers, buffers (e.g., blocking buffers or lysis buffers), etc. may also be included. The relative amounts of a variety of reagents can be changed to provide concentrates of reagent solutions, which provides user flexibility, space saving, reagent saving, etc. These reagents may also be provided in the form of a dry powder, usually lyophilized, including excipients which, when dissolved, provide a solution of the reagent having the appropriate concentration.

[0225] In one aspect, the disclosure provides use of the antibodies or antigen-binding portions thereof, bispecific or multispecific molecules, nucleic acid molecules, vectors, cells and / or compositions in the preparation of a medicament or kit for diagnosing, treating or preventing a disease associated with MASP-2-dependent complement activation.

[0226] In addition, the antibodies disclosed herein can also be used in immunoassays, purification methods, and other methods using immunoglobulins or fragments thereof. Such uses are well known in the art.

[0227] Accordingly, the present disclosure also provides compositions comprising the anti-MASP-2 antibodies or fragments thereof of the present disclosure, which are conveniently combined with a pharmaceutically acceptable carrier, diluent or excipient, as is conventional in the art.

[0228] The term "pharmaceutical composition" used in the present disclosure refers to a variety of preparations. The formulation containing a therapeutically effective amount of a multivalent antibody is in the form of a sterile liquid solution, liquid suspension or lyophilized form, optionally containing a stabilizer or excipient.

[0229] It will be understood that the therapeutic agents according to the described embodiments will be administered with suitable pharmaceutically acceptable carriers, excipients, and other agents incorporated into formulations to provide improved transfer, delivery, tolerance, etc. A large number of suitable formulations can be found in the pharmacopoeia known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences (15th edition, Mack Publishing Company, Easton, Pa. (1975)), especially Chapter 87 of Blaug, Seymour therein. These formulations include, for example, powders, pastes, ointments, gels, waxes, oils, lipids, lipid (cationic or anionic) carriers (e.g., Lipofectin TM ), DNA conjugates, anhydrous slurries, oil-in-water and water-in-oil emulsions, emulsion polyethylene glycol (polyethylene glycol of various molecular weights), semisolid gels, and semisolid mixtures containing polyethylene glycol. Any of the foregoing mixtures may be suitable for treatment or therapy according to the present disclosure, provided that the active ingredient in the formulation is not inactivated by the formulation and that the formulation is physiologically compatible and tolerates the route of administration.

[0230] In another embodiment, antibodies against MASP-2 can be used in methods known in the art relating to the localization and / or quantification of MASP-2 (e.g., for determining MASP-2 and / or the level of MASP-2 in an appropriate physiological sample, for diagnostic methods, for protein imaging, etc.). In a given embodiment, an antibody specific for MASP-2, or a derivative, fragment, analog or homolog thereof, comprising an antigen binding domain derived from the antibody, is used as a pharmaceutically active compound (hereinafter referred to as a "therapeutic agent").

[0231] In another embodiment, the MASP-2 polypeptide can be isolated by standard techniques such as immunoaffinity, chromatography, or immunoprecipitation using antibodies specific for MASP-2. Antibodies (or fragments thereof) directed against MASP-2 proteins can be used to detect the protein in biological samples. In some embodiments, MASP-2 can be detected in biological samples as part of a clinical testing process, for example, to determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin / biotin and avidin / biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazineaminofluorescein, dansyl chloride, or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive materials include 125 I. 131 I. 35 S or 3 H.

[0232] In another embodiment, the antibodies according to the present disclosure can be used as reagents for detecting the presence of MASP-2 or a protein fragment thereof in a sample. In some embodiments, the antibody comprises a detectable label. The antibody is a polyclonal antibody, or more preferably a monoclonal antibody. The use of intact antibodies or fragments thereof (e.g., Fab, scFv or F(ab') 2). The term "labeling" about an antibody is intended to include direct labeling of the antibody by coupling (i.e., physically connecting) a detectable substance to the antibody, and indirect labeling of the antibody by reacting with another reagent of direct labeling. Examples of indirect labeling include detecting the first antibody using a fluorescently labeled second antibody, and terminally labeling the antibody with biotin so that it can be detected with fluorescently labeled streptavidin. The term "biological sample" is intended to include tissues, cells, and biological fluids separated from a subject, as well as tissues, cells, and fluids present in the subject. Therefore, the term "biological sample" used includes blood and fractions or components in the blood, including serum, plasma, or lymph. In other words, the detection method of the embodiment can be used to detect analyte mRNA, protein, or genomic DNA in a biological sample in vitro and in vivo. For example, analyte mRNA in vitro detection techniques include Northern hybridization and in situ hybridization. Analyte protein in vitro detection techniques include enzyme-linked immunosorbent assay (ELISA), Western blotting, immunoprecipitation, and immunofluorescence. Analyte genomic DNA in vitro detection techniques include Southern hybridization. Procedures for performing immunoassays are described, for example, in "ELISA: Theory and Practice: Methods in Molecular Biology", Vol. 42, JR Crowther (ed.) Human Press, Totowa, NJ, 1995; "Immunoassay", E. Diamandis and T. Christopoulus, Academic Press, Inc., San Diego, Calif., 1996; and "Practice and Theory of Enzyme Immunoassays", P. Tijssen, Elsevier Science Publishers, Amsterdam, 1985. In addition, in vivo detection techniques for analyte proteins include introducing into a subject a labeled anti-analyte protein antibody. For example, the antibody can be labeled with a radioactive label, and the presence and location of the radioactive label in the subject can then be detected by standard imaging techniques.

[0233] The antibodies and derivatives, fragments, analogs and homologues described herein can be incorporated into pharmaceutical compositions suitable for administration. The principles and considerations involved in preparing such compositions and guidelines for selecting components are well known in the art, for example, see Remington's Pharmaceutical Sciences: The Science And Practice Of Pharmacy 19th Edition (Alfonso R. Gennaro et al., ed.) Mack Pub. Co., Easton, Pa.: 1995; Drug Absorption Enhancement: Concepts, Possibilities, Limitations, And Trends, Harwood Academic Publishers, Langhorne, Pa., 1994; and Peptide And Protein Drug Delivery (Advances In Parenteral Sciences, Vol. 4), 1991, M. Dekker, New York.

[0234] Such compositions generally comprise an antibody and a pharmaceutically acceptable carrier. When an antibody fragment is used, the smallest inhibitory fragment that specifically binds to the target protein binding domain may be preferred. For example, based on the variable region sequence of the antibody, a peptide molecule that retains the ability to bind to the target protein sequence can be designed. Such peptides can be chemically synthesized and / or produced by recombinant DNA technology (see, e.g., Marasco et al., Proc. Natl. Acad. Sci. USA, 90: 7889-7893 (1993)).

[0235] As used herein, the term "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc., that are compatible with drug administration. Suitable pharmaceutically acceptable carriers are described in the latest edition of Remington's Pharmaceutical Sciences, a standard reference work in this field, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous carriers, such as immobilized oils, may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except for any conventional media or agents that are incompatible with the antibody, its use in the composition is contemplated.

[0236] The pharmaceutical composition of the embodiment is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, such as intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration. Solutions or suspensions for parenteral, intradermal, or subcutaneous administration may include the following components: sterile diluents for injection such as water, saline, fixed oils, polyethylene glycols, glycerol, propylene glycol, or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates, or phosphates, and agents for regulating osmotic pressure such as sodium chloride or dextrose. pH can be adjusted with an acid or a base, such as hydrochloric acid or sodium hydroxide. Parenteral preparations can be packaged in ampoules, disposable syringes, or glass or plastic multidose vials.

[0237] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable pharmaceutically acceptable carriers include physiological saline, bacteriostatic water, Cremophor EL TM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that it is easy to inject. It must be stable under manufacturing and storage conditions and must be able to prevent the contamination of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyols (for example, glycerol, propylene glycol and liquid polyethylene glycol, etc.), and a suitable mixture thereof. For example, by using a coating such as lecithin, maintaining the required particle size in the case of a dispersion, and using a surfactant, suitable fluidity can be maintained. The prevention of microbial action can be achieved by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, etc. In many cases, it will be preferred to include isotonic agents in the composition, such as sugars, polyols (such as mannitol, sorbitol), sodium chloride. The extended absorption of the composition for injection can be achieved by including an agent such as aluminum monostearate and gelatin that delays absorption in the composition.

[0238] As required, sterile injection solutions can be prepared by incorporating the antibody in the required amount into a suitable solvent having one or a combination of the above-listed components (as required), followed by filtration sterilization. Generally speaking, dispersions are prepared by incorporating the antibody into a sterile carrier containing an alkaline dispersion medium and the other required components listed above. With regard to sterile powders for the preparation of sterile injection solutions, the preparation method is vacuum drying and freeze drying to obtain a powder comprising the active ingredient and any additional desired components from a sterile filtered solution of the aforementioned components.

[0239] For administration by inhalation, the compounds are delivered in the form of an aerosol spray from a pressured container or dispenser which contains a suitable propellant, such as a gas such as carbon dioxide, or a nebulizer.

[0240] Administration can also be systemically administered via mucosal or transdermal means. For mucosal or transdermal administration, a penetrant suitable for permeation of the barrier is used in the formulation. Such penetrants are generally known in the art and include detergents, bile salts and fusidic acid derivatives such as those used for mucosal administration. Mucosal administration can be achieved by using nasal sprays or suppositories. For transdermal administration, one or more of the antibodies can be formulated into an ointment, an ointment, a gel or a cream as is generally known in the art.

[0241] The compounds may also be prepared for rectal delivery in the form of suppositories (eg, with conventional suppository bases such as cocoa butter or other glycerides) or retention enemas.

[0242] In one embodiment, the antibody can be prepared with a carrier that prevents it from being rapidly eliminated by the body, such as a sustained / controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene-vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparing such formulations will be apparent to those skilled in the art.

[0243] It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and consistency of dosage. As used herein, dosage unit form refers to physically separable units suitable as unit dosages for the subject to be treated; each unit contains a predetermined amount of one or more of the antibodies calculated to produce the desired therapeutic effect in combination with the required pharmaceutical carrier. The specifications for the dosage unit form of the embodiments are dictated by and directly dependent upon the unique characteristics of the antibodies and the specific therapeutic effect to be achieved, and the limitations inherent in the art of formulating such antibodies for treating individuals.

[0244] The pharmaceutical compositions can be placed in a container, pack, or dispenser together with instructions for administration.

[0245] The formulations described herein may also include more than one of the antibodies, preferably those with complementary activities but without negative effects on each other, depending on the specific condition to be treated. Alternatively or in addition, the composition may, for example, include agents that enhance its function, such as cytotoxic agents, cytokines, chemotherapeutic agents, or growth inhibitors. Such molecules are appropriately combined in an amount effective for the intended purpose. For example, they may be combined in a kit or in use.

[0246] In one embodiment, one or more of the antibodies may be administered in a combined therapy, i.e., in combination with other agents such as therapeutic agents (which can be used to treat pathological conditions or disorders, such as various forms of autoimmune diseases and inflammatory diseases). The term "combined" herein refers to administering the agents substantially synchronously, simultaneously or sequentially. If administered sequentially, the first of the two compounds is still preferably detected at an effective concentration at the treatment site when the second compound is administered. In one case, "combined" may also be the simultaneous inclusion of the disclosed antibodies and other therapeutic agents in a kit.

[0247] For example, a combination therapy may comprise one or more antibodies described herein co-formulated and / or co-administered with one or more additional therapeutic agents (e.g., one or more cytokine and growth factor inhibitors, immunosuppressants, anti-inflammatory agents, metabolic inhibitors, enzyme inhibitors, and / or cytotoxins or cytostatic agents, as described in more detail below). Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thereby avoiding possible toxicities or complications associated with various monotherapies.

[0248] The experiments disclosed herein prove that the various monoclonal antibodies and variants thereof provided by the disclosure that can bind to MASP-2 with high affinity and inhibit lectin-mediated complement activation can specifically recognize the enzymatic domains CCP1-CCP2-SP of human, monkey, mouse and rat MASP-2, and can effectively inhibit the generation of C4b and C3 convertase C4b2b under low concentration conditions, and its blocking activity is 50-200 times higher than that of known antibodies. The results of pre-toxicological studies conducted in rhesus monkeys show that animals tolerate high doses of new MASP-2 antibodies well without any toxic reactions, and its efficacy and safety are better than similar antibodies.

[0249] For purposes of clarity and concise description, features are described herein as part of the same or separate embodiments, however, it will be understood that the scope of the present disclosure may include embodiments having a combination of all or some of the described features.

[0250] Example

[0251] Example 1. Detection of MASP-2 activity in the lectin pathway

[0252] After MBL in plasma recognizes repeated N-galactose or mannose structures, it can sequentially activate serine proteases MASP-1 and MASP-2. Activated MASP-2 cleaves complement components C4 and C2, and the resulting C4b is deposited on the well plate and can be recognized by anti-C4b antibodies. C3 convertase C4b2b, formed by the cleavage products of C4 and C2, cleaves complement component C3, and the resulting C3b is deposited on the well plate and can be recognized by anti-C3b antibodies. Therefore, by detecting the production of C4b or C3b in the lectin activation reaction, the inhibitory activity of MASP-2 antibodies on MASP-2 in serum can be displayed.

[0253] C4 cleavage activity assay

[0254] 20 μg / ml mannan (Sigma, M7504) was added to the ELISA plate and coated overnight at 4°C. After blocking with 2% BSA at room temperature for 60 minutes, 0.5% diluted human serum was added and incubated overnight at 4°C. After washing, 50 μl / well of MASP-2 antibody of different concentrations (starting concentration was 30 μg / ml, 5-fold gradient dilution, 9 gradients in total) was added, and after pre-incubation at 4°C for 30 minutes, 50 μl of 2 μg / ml human C4 protein (CompTech, #A105) was added and reacted at 37°C for 90 minutes. After washing, add 1:1000 diluted Biotin-labeled human C4b polyclonal antibody (ASSAYPRO, #11223-05021) and incubate at room temperature for 90 minutes. Add 1:5000 peroxidase-streptavidin (Jackson ImmunoResearch 016-030-084) and incubate at room temperature for 60 minutes. Add TMB color development solution and color at 37°C in the dark for about 10 minutes. Terminate the reaction and read the OD value.

[0255] C3 convertase activity assay

[0256] 20 μg / ml mannan was added to the ELISA plate and coated overnight at 4°C. After blocking with 2% BSA at room temperature for 60 minutes, MASP-2 antibody diluted with 0.5% human serum was added (initial concentration was 60 μg / ml, 5-fold gradient dilution, 9 gradient dilutions in total), reacted at 37°C for 90 minutes, 1:500 rabbit anti-human C3 / C3b / C3c polyclonal antibody (Proteintech, #21337-1-AP) was added, incubated at room temperature for 60 minutes, 1:5000 HRP-labeled goat anti-rabbit secondary antibody (Jackson ImmunoResearch111-035-144) was added, incubated at room temperature for 60 minutes, TMB color development solution was added, color was developed at 37°C in the dark for about 10 minutes, the reaction was terminated and the OD value was read.

[0257] Example 2. Preparation of MASP-2 recombinant protein

[0258] The sequence encoding the human MASP-2 enzymatic domain CCP1-CCP2-SP was cloned into a eukaryotic expression plasmid, and the N-terminal his6 tag was fused for easy purification. After transfection of HEK293E cells and culture for 7 days, the culture supernatant was collected and purified by nickel chelate column to obtain human MASP-2D4-6 recombinant protein (SEQ ID NO.1). Similarly, cynomolgus monkey MASP-2D4-6 (SEQ ID NO.2), mouse MASP-2D4-6 (SEQ ID NO.3) and rat MASP-2D4-6 (SEQ ID NO.4) recombinant proteins were obtained. Figure 1 Shown are the SDS-PAGE results of human, cynomolgus monkey, mouse and rat MASP-2D4-6 recombinant proteins.

[0259] Example 3. Animal immunization and preparation of hybridoma antibodies

[0260] Six-week-old BALB / c female mice were selected and immunized subcutaneously or in the footpad using human or cynomolgus monkey MASP-2D4-6 recombinant protein mixed with Freund's adjuvant in equal volumes. Two weeks after the first immunization, booster immunization was performed, and shock immunization was performed when the serum titer of the immunized mice reached 1:100,000. Three days later, the spleen and peripheral lymph nodes of the mice were taken to isolate B cells.

[0261] Take appropriate amount of B cells and SP2 / 0 cells for fusion, screen in DMEM complete medium containing HAT, and place hybridoma cells in 5% CO 2 Cultured at 37°C for 5-7 days. Human or cynomolgus monkey MASP-2D4-6 recombinant protein (1 μg / mL) was coated and hybridoma culture supernatant was tested by ELISA. A total of 778 monoclonal antibodies could cross-recognize human or cynomolgus monkey MASP-2. The supernatant was harvested and tested by C4 cleavage activity. 61 clones had significant MASP-2 inhibitory activity (such as Figure 2 shown).

[0262] According to the results of hybridoma screening, RNA of candidate hybridoma monoclonal cells was extracted by TRNzol lysis method, and then reverse transcribed to synthesize single-stranded cDNA, which was used as a template to amplify the variable region sequence of the antibody in the hybridoma monoclonal cells.

[0263] The variable region sequences of the MASP-2 antibody are as follows:

[0264] Table 1. Variable region sequences of MASP-2 mouse antibodies

[0265]

[0266]

[0267] Clone: ​​1B2

[0268] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.5, its encoding nucleic acid is shown in SEQ ID NO.6, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 8 and 9 respectively.

[0269] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFSGDKLEWMG YISYSGSTSYNPSLKS RIS------------FR3------------->CDR3<---FR4--->ITRDTSKNQFFLHLSSVTTEDTATYFCAR YWGDY WGHGTTLTVSS

[0270] Nucleic acid sequence

[0271] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTTCAGGAGACAAACTGGAGTGGATGGGCTACATAAGCTACAGTGGTTCC ACTAGCTATAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCACTTGAGTTCTTGTGACTACTGAGGACACAGCCACATATTTCTGTGCAAGATATTGGGGCGACTACTGGGGCCACGGCACCACTCTCACAGTCTCCTCA

[0272] The amino acid sequence of the light chain VK is shown in SEQ ID NO.10, the encoding nucleic acid is shown in SEQ ID NO.11, and the CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.12, 13 and 14 respectively.

[0273] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DSVMTQSQKFMSTSVGDRVSVTC KASHNVGTNVA WFQQKPGQSPKGLIYSASYRSS GVPDRFTGSGSGT-----FR3---------->CDR3<---FR4--->DFTLTISNVQSEDLAEYFC LQYNSNPLT FGGGTKLEVK

[0274] Nucleic acid sequence

[0275] GACAGTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCACAATGTGGGTACTAATGTAGCCTGGTTTCAACAGAAACCAGGACAATCTCCTAAAGGACTGATTTACTCGGCATCCTACC GGTCCAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGACTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTATTTCTGTCTACAATATAACAGCAATCCTCTGACGTTCGGTGGAGGCACCAAGCTGGAGGTCAAG

[0276] Clone: ​​3B9

[0277] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.15, the encoding nucleic acid is shown in SEQ ID NO.16, and the CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 18 respectively.

[0278] <-------------FR1---------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVRPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPSLKS RLS------------FR3------------->CDR3<---FR4--->ITRDTSKNQFFLQLNSVSSADTATYYCAI ISTAVDFDY WGQGTTLTVSS

[0279] Nucleic acid sequence

[0280] GATGTACAACTTCAGGAGTCGGGACCTGGCCTGGTGAGACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGTTACAGTGGTCGCACTAGT TACAACCCCTCTCAAAAGTCGACTCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGAGTTCTGCGGACACAGCCACGTATTACTGTGCAATTATTTCTACGGCAGTAGACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA

[0281] The amino acid sequence of the light chain VK is shown in SEQ ID NO.19, its encoding nucleic acid is shown in SEQ ID NO.20, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.21, 22 and 23 respectively.

[0282] <----------FR1-------->CDR1<-----FR2----->CDR2<------------QTVLTQSPAIMSASLGERVTMTC TASSSVSSSYLL WYQQKPGSSPKLWIY STSKLAS GVPVRFSGSGSG-----FR3----------->CDR3<---FR4--->TSYSLTISSMEADDAATYYC LQYNRSPLT FGAGTKLELK

[0283] Nucleic acid sequence

[0284] CAAAcTGTTCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCTAGGGGAACGGGTCACCATGACCTGCACTGCCAGCTCAAGTGTAAGTTCCAGTTACTTGCTCTGGTACCAGCAGAAGCCAGGATCCTCCCCCAAACTCTGGATTTATAGCACATCCAAG TTGGCTTCTGGAGTCCCAGTTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGACGATGCTGCCACTTATTACTGCCTCCAGTATAATCGTTCCCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0285] Clone: ​​4F8

[0286] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.24, its encoding nucleic acid is shown in SEQ ID NO.25, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.26, 27 and 28 respectively.

[0287] <-------------FR1----------->CDR1<----FR2----->CDR2<--EVQLQESGPGLVKPSQSLSLACTVTGYSI TSDYAWH WIRQFPGNKLEWMG FISYSGRTNYNPPLTS RIS------------FR3------------->CDR3<---FR4--->

[0288] ITRDTSKNQFFLQLNSVTTEDTATYYCAR SYDYWFGY WGQGTLVTVSA

[0289] Nucleic acid sequence

[0290] GAGGTCCAGCTGCAGGAGTCGGGACCTGGCCTGGTGAAACCTTTCCAGTCTCTGTCCCTCGCCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGCACTGGATCCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTTCATAAGCTACAGTGGTAGAACTA ACTACAACCCACCTCTCACAAGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGATCATATGATTACTGGTTTGGTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA

[0291] The amino acid sequence of the light chain VK is shown in SEQ ID NO.29, its encoding nucleic acid is shown in SEQ ID NO.30, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.31, 32 and 33 respectively.

[0292] <----------FR1-------->CDR1<-----FR2----->CDR2<-------NVLMTQTPLSLPVSLGDQASISC RSSQSLVHSNGNTYLE WYMQKPGQSPKLLIY TVSRRFS GVPDRFSG-----FR3----------->CDR3<---FR4--->

[0293] SGSGTDFTLKISRVEAEDLGVYYC FQGSHVPVT FGAGTKLELK

[0294] Nucleic acid sequence

[0295] AATGTTTTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGTCTTGTACATAGTAATGGAAACACCTATTTAGAATGGTACATGCAGAAACCAGGCCAGTCTCCAAAGCTCCTGATCTACACAGTT TCCAGACGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGATGGAGGCTGAGGATCTGGGAGTTTATTGCTTTCAAGGTTCACATGTTCCGGTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0296] Clone: ​​5J11

[0297] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.34, its encoding nucleic acid is shown in SEQ ID NO.35, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.36, 37 and 38 respectively.

[0298] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWK WIRQFPGNKLEWMG YISYSGSTSYNPSLKG RIS------------FR3------------->CDR3<---FR4--->

[0299] ITRDTSKNQFFLHLQSVTIEDTATYYCAI LGYFAMDY WGQGTPVTVSS

[0300] Nucleic acid sequence

[0301] GATGTACAACTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAAGTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGTTACAGTGGCAGCACTA GCTACAACCCTTCTCTCAAAGGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTTTTCCTGCACTTGCAATCTGTGACTATTGAGGACACAGCCACCTATTACTGTGCAATACTAGGTTACTTTGCTATGGACTACTGGGGGCAAGGAACCCCAGTCACCGTCTCCTCA

[0302] The amino acid sequence of the light chain VK is shown in SEQ ID NO.39, its encoding nucleic acid is shown in SEQ ID NO.40, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.41, 42 and 43 respectively.

[0303] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFMSTSVGDRVSITC KASQNVRVDVA WYQQKSGQSPKSLIY LASNRHT GVPDRFTGSGSGT-----FR3---------->CDR3<---FR4--->

[0304] DFTLTISNVQSEDLADYFC LQHWSYPLT FGGGTKLEIK

[0305] Nucleic acid sequence

[0306] GACATTTGTGATGACCCAGTCTCAAAAATTCATGTCCACTTCAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTTCGTGTTGACGTTGCCTGGTATCAACAGAAATCAGGGCAGTCTCCTAAATCACTGATTTACTTGGCATCCAACC GGCACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTAGCAATGTGCAATCTGAAGACCTGGCAGATTATTTCTGTCTACAACATTGGAGTTATCCTCTGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA

[0307] Clone: ​​9P13

[0308] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.44, its encoding nucleic acid is shown in SEQ ID NO.45, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 46 and 47, respectively.

[0309] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNTLEWMG YISYSGRTSYNPSLIG RIS------------FR3------------->CDR3<---FR4--->

[0310] ITRDTTKNQFFLQLNSVSTEDTATYYCAK LAGGHGMDY WGQGTSVTVSS

[0311] Nucleic acid sequence

[0312] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACACATTGGAGTGGATGGGCTATATAAGTTATAGTGGTAGGACTAGC TACAACCCATCTCTGATAGGTCGAATCTCAATCACTCGAGACACAACCAAGAACCAGTTCTTTCTGCAGTTGAATTCTGTGTCTACTGAGGACACAGCCACATATTACTGTGCAAAATTGGCCGGAGGACATGGGATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0313] The amino acid sequence of the light chain VK is shown in SEQ ID NO.48, its encoding nucleic acid is shown in SEQ ID NO.49, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.50, 51 and 52 respectively.

[0314] <----------FR1-------->CDR1<-----FR2----->CDR2<------------NIVMTQSQQFMSTSVGDRVSVTC KASQNVDTNVA WYQQKPGQSPKALIY SASYRYS GVPDRFTGSGSGT-----FR3---------->CDR3<---FR4--->

[0315] DFTLTISNVQSEDLAEYFC LQYNSYLT FGSGTKLELK

[0316] Nucleic acid sequence

[0317] AAATTGTGATGACCCAGTCTCAACAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAGAAACCAGGACAATCTCCTAAAGCACTGATTTACTCGGCATCCTAC CGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAATATTTCTGTCTGCAATATAATAGTTACCTCACGTTCGGTTCTGGGACCAAGCTGGAGCTGAAA

[0318] Clone: ​​10H9

[0319] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.53, its encoding nucleic acid is shown in SEQ ID NO.54, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 18 respectively.

[0320] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVRPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPSLKS RLS------------FR3------------->CDR3<---FR4--->

[0321] VTRDTSKNQFFLQLNSVNAADTATYYCAI ISTAVDFDY WGQGTTLTVSS

[0322] Nucleic acid sequence

[0323] GATGTACAACTTCAGGAGTCGGGACCTGGCCTGGTGAGACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGTTACAGTGGTCGCACTAGC TACAACCCCTCTCTCAAGAGTCGACTCTCTGTCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGAATGCTGCGGACACAGCCACGTATTACTGTGCAATTATTTCTACGGCAGTAGACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA

[0324] The amino acid sequence of the light chain VK is shown in SEQ ID NO.55, its encoding nucleic acid is shown in SEQ ID NO.56, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.21, 57 and 23, respectively.

[0325] <----------FR1-------->CDR1<-----FR2----->CDR2<------------QIVLTQSPAIMSASLGERVTMTC TASSSVSSSYLL WYQQKPGSSPKLWIY SSSKLAS GVPVRFSGSGSG-----FR3----------->CDR3<---FR4-->

[0326] TSYSLTISSMEAEDAATYYC LQYNRSPLT FGAGTKLELK

[0327] Nucleic acid sequence

[0328] CAAATTGTTCTCACCCAGTCTCCAGCAATCATGTCTGCATCTCTAGGGGAACGGGTCACCATGACCTGCACTGCCAGCTCAAGTGTAAGTTCCAGTTACTTGCTCTGGTACCAGCAGAAGCCAGGATCCCTCCCCCAAACTCTGGATTTATAGCTCATCCAAG TTGGCTTCTGGAGTCCCAGTTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCTCTCACAATCAGCAGCATGGAGGCTGAAGATGCTGCCACTTATTACTGCCTCCAGTATAATCGTTCCCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0329] Clone: ​​10L3

[0330] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.58, its encoding nucleic acid is shown in SEQ ID NO.59, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 60 respectively.

[0331] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPSLKS RIS------------FR3------------->CDR3<---FR4--->

[0332] ITRDTSKNQFFLQLNSVTTEDTATYYCAK LGGGHGMDY WGQGTSVTVSS

[0333] Nucleic acid sequence

[0334] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAATTGGAGTGGATGGGCTACATAAGTTATAGTGGTAGGACTAGC TACAACCCATCTCTGAAAAGTCGAATCTCAATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAAATTGGGCGGAGGACATGGGATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0335] The amino acid sequence of the light chain VK is shown in SEQ ID NO.61, its encoding nucleic acid is shown in SEQ ID NO.62, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.50, 51 and 52 respectively.

[0336] <----------FR1-------->CDR1<-----FR2----->CDR2<-------------IVMTQSQKFMSTSVGDRVSVTC KASQNVDTNVA WYQQKPGQSPKALIY SASYRYS GVPDRFTGSGSGTD----FR3---------->CDR3<---FR4--->

[0337] FTLTISNVQSEDLAEYFC LQYNSYLT FGSGTKLELK

[0338] Nucleic acid sequence

[0339] ACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCTAAAGCGCTGATTTACTCGGCATCCTAC CGTTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTATTTCTGTCTGCAATATAACAGCTACCTCACGTTCGGTTCTGGGACCAAGCTGGAGCTGAAA

[0340] Clone: ​​11F8

[0341] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.63, the encoding nucleic acid thereof is shown in SEQ ID NO.64, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NO.65, 66 and 67, respectively. <-------------FR1----------->CDR1<----FR2----->CDR2<-QVTLKESGPGILKPSQTLILTCSFSGFSL NTSGMGVG WIRQPSGKGLEWLA HIWWD DDKDYNPSLKS QV------------FR3-------------->CDR3<---FR4--->

[0342] TISKDTTSNQVFLKITSVDTADGATYYCVR DTTFLDAMDY WGQGTSVTVSS

[0343] Nucleic acid sequence

[0344] CAAGTTACTCTAAAAGAGTCTGGCCCTGGGATATTGAAGCCCTCACAGACCCTCATTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAACACTTCTGGTATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGGATGATGATAAGGAC TATAACCCATCCCTGAAGAGCCAAGTCACAATCTCCAAGGATACCTCCACAAACCAGGTGTTCCTCAAGATCACCAGTGTGGACACTGCAGATGGTGCCACTTACTACTGTGTTCGAGATACTACGTTTTTAGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0345] The amino acid sequence of the light chain VK is shown in SEQ ID NO.68, its encoding nucleic acid is shown in SEQ ID NO.69, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.70, 71 and 72 respectively.

[0346] <----------FR1-------->CDR1<-----FR2----->CDR2<-------DVLMTQTPLSLPVSLGDQASISC RSSQSIVHRNGNTFLE WYLQKPGQSPKLLIY RVSNRFS GVPDRFSG-----FR3-------------->CDR3<---FR4-->

[0347] SGSGTDFTLKISRVEAEDLGVYYC FQGSHVPFT FGSGTKLEIK

[0348] Nucleic acid sequence

[0349] GATGTTTTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGGTCTAGTCAGAGCATTGTACATCGTAATGGAAACACCTTTTTAGAATGGTACCTGCAGAAACCAGGCCAGTCTCCAAAGCTCCTGATCTACAGGGTT TCCAACCGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGATGGAGGCTGAGGATCTGGGAGTTTATTACTGCTTTCAAGGTTCACATGTTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA

[0350] Clone: ​​11G8

[0351] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.73, its encoding nucleic acid is shown in SEQ ID NO.74, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 47, respectively.

[0352] <-------------FR1---------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQLPGNKLEWVG YISYSGRTSYNPSLKS RIS------------FR3------------->CDR3<---FR4--->

[0353] ITRDTTKNQFFLQLNSVTTEDTATYYCAK LAGGHGMDY WGQGTSVTVSS

[0354] Nucleic acid sequence

[0355] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTACCAGGAAACAAATTGGAGTGGGTGGGCTATATAAGTTATAGTGGTAGGACTAGC TACAACCCATCTCTGAAAAGTCGAATCTCAATCACTCGAGACACAACCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAAGTTGGCCGGGGGACATGGGATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0356] The amino acid sequence of the light chain VK is shown in SEQ ID NO.75, its encoding nucleic acid is shown in SEQ ID NO.76, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.50, 51 and 52 respectively.

[0357] <----------FR1-------->CDR1<-----FR2----->CDR2<-------------NIVMTQSQKFMSTSVGDRVSVTC KASQNVDTNVA WYQQKPGQSPKALIH SASYRYS GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0358] DFILTISNVQSEDLAEYFC LQYNSYLT FGSGTKLELK

[0359] Nucleic acid sequence

[0360] aACaTTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCTAAAGCACTGATTCACTCGGCATCCTAC CGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCATTCTCACCATCAGTAATGTGCAGTCTGAAGACTTGGCAGAGTATTTCTGTCTGCAATATAACAGCTACCTCACGTTCGGTTCTGGGACCAAGCTGGAGCTGAAA

[0361] Clone: ​​14K22

[0362] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.77, its encoding nucleic acid is shown in SEQ ID NO.78, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 79, respectively.

[0363] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPSLKS RIS------------FR3------------->CDR3<---FR4--->ITRDTSKNQFFLHLNSVTTEDTATYFCAV LGLYFDY WGQGTPLTVSS

[0364] Nucleic acid sequence

[0365] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGCTACAGTGGTAGAACT AGCTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCACTTGAATTCTGTGACTACTGAGGACACAGCCACATATTTCTGTGCAGTACTGGGACTCTACTTTGACTACTGGGGCCAAGGCACCCCTCTCACAGTCTCCTCA

[0366] The amino acid sequence of the light chain VK is shown in SEQ ID NO.80, the encoding nucleic acid thereof is shown in SEQ ID NO.81, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NO.50, 51 and 82, respectively. <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFMSTSVGDRVSVTC KASQNVDTNVA WYQQKPGQSPKALIY S ASYRYS GVPDRFTGSGSGT

[0367] ----FR3----------->CDR3<---FR4--->

[0368] DFTLTISNVQSEDLAEYFC HQFNSNPLT FGAGTKLELK

[0369] Nucleic acid sequence

[0370] GACATTTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAAGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCTAAAGCACTGATTTACTCGGCATCCTACC GGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAATATTTCTGTCACCAATTTAACAGCAATCCTTCCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0371] Clone: ​​16A15

[0372] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.83, its encoding nucleic acid is shown in SEQ ID NO.84, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.36, 85 and 86, respectively.

[0373] <-------------FR1---------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWK WIRQFPGNKLEWMG YISYSGSTSYHPSLKG RIS------------FR3------------->CDR3<---FR4--->

[0374] ITRDTSKNQFFLQLNSVTADDTATYYCAI LGYYAMDY WGQGTSVTVSS

[0375] Nucleic acid sequence

[0376] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAAATGGATCCGGCAGTTTCCAGGAAACAAACTGGAATGGATGGGCTACATAAGCTACAGTGGTTCCACTA GCTATCACCCATCTCTCAAAGGTCGAATCTCCATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTGCTGACGACACAGCCACATATTACTGTGCAATATTAGGTTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0377] The amino acid sequence of the light chain VK is shown in SEQ ID NO.87, its encoding nucleic acid is shown in SEQ ID NO.88, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.41, 42 and 43, respectively.

[0378] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFMSTSVGDRVSITC KASQNVRVDVA WYQQKSGQSPRALIY LASNRHT GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0379] DFTLTITNVQSEDLADYFC LQHWSYPLT FGGGTKLEIK

[0380] Nucleic acid sequence

[0381] GACATTTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGTATCACCTGCAAGGCCAGTCAGAATGTTCGTGTTGACGTAGCCTGGTATCAACAGAAATCAGGGCAGTCTCCTAGAGCACTGATTTACTTGGCATCCAACC GGCACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTACCAATGTGCAATCTGAGGACCTGGCAGATTATTTCTGTCTACAACATTGGAGTTATCCTCTGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA

[0382] Clone: ​​16A17

[0383] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.83, its encoding nucleic acid is shown in SEQ ID NO.84, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.36, 85 and 86, respectively.

[0384] <-------------FR1---------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWK WIRQFPGNKLEWMG YISYSGSTSYHPSLKG RIS------------FR3------------->CDR3<---FR4--->

[0385] ITRDTSKNQFFLQLNSVTADDTATYYCAI LGYYAMDY WGQGTSVTVSS

[0386] Nucleic acid sequence

[0387] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAAATGGATCCGGCAGTTTCCAGGAAACAAACTGGAATGGATGGGCTACATAAGCTACAGTGGTTCCACTA GCTATCACCCATCTCTCAAAGGTCGAATCTCCATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTGCTGACGACACAGCCACATATTACTGTGCAATATTAGGTTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0388] The amino acid sequence of the light chain VK is shown in SEQ ID NO.89, its encoding nucleic acid is shown in SEQ ID NO.90, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.41, 42 and 91, respectively.

[0389] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFMSTSVGDRVSITC KASQNVRVDVA WYQQKSGQSPRALIY LASNRHT GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0390] DFTLTITNVQSEDLADYFC PQHWSYPLT FGGGTKLEIK

[0391] Nucleic acid sequence

[0392] GACATTTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGTATCACCTGCAAGGCCAGTCAGAATGTTCGTGTTGACGTAGCCTGGTATCAACAGAAATCAGGGCAGTCTCCTAGAGCACTGATTTACTTGGCATCCAACC GGCACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTACCAATGTGCAATCTGAGGACCTGGCAGATTATTTCTGTCcACAACATTGGAGTTATCCTCTGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA

[0393] Clone: ​​20J6

[0394] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.92, its encoding nucleic acid is shown in SEQ ID NO.93, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.94, 95 and 96, respectively.

[0395] <-------------FR1----------->CDR1<----FR2----->CDR2<--KVKLVESGGDLVKPGGSLKISCTASGFTF SNYAMS WIRQTPDKRLEWVA TVSSGGPYTYYPDSVKG RFT------------FR3------------->CDR3<---FR4--->

[0396] ISRDNAENTLYLQMSSLRSDDTAIYYCAR QIYSDYDVTY WGQGTLVTVSA

[0397] Nucleic acid sequence

[0398] aAAGTGAAGCTGGTGGAGTCTGGGGGAGACTTAGTGAAGCCTGGAGGGTCCCTGAAAATCTCCTGTACAGCCTCTGGATTCACTTTCAGTAACTATGCCATGTCTTGGATTCGCCAGACTCCGGACAAGAGGCTGGAGTGGGTCGCAACCGTTAGTAGTGGTGGTCCTTACACCTACT ATCCAGACAGTGTGAAGGGGCGATTCACCATCTCCAGAGACAATGCCGAAAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGATGACACGGCCATATATTACTGTGCAAGGCAGATCTACTCTGATTACGACGTCACTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA

[0399] The amino acid sequence of the light chain VK is shown in SEQ ID NO.97, its encoding nucleic acid is shown in SEQ ID NO.98, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.99, 100 and 101 respectively.

[0400] <----------FR1-------->CDR1<-----FR2----->CDR2<-------------QIVLSQSPAILSASPGETVTMTC RASSSVSFMY WLQQKPGSSPQPWIY ATSNLAS GVPARFSGSGSGTS----FR3---------->CDR3<---FR4--->

[0401] YSLTITRVEAEDAATYYC QQWSFRPWT FGGGTKLEIK

[0402] Nucleic acid sequence

[0403] CAAATTGTTCTTCCCAGTCTCCAGCAATCCTGTCTGCATCTCCAGGGGAGACGGTCACAATGACTTGCAGGCCAGTTCAAGTGTAAGTTTCATGTACTGGCTCCAGCAGAAGCCAGGTTCCTCCCCCCAACCCTGGATTTATGCCACATCCAACCTG GCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCCCTCACAATCACCAGAGTGGAGGCTGAAGATGCTGCCACTTATTACTGTCAACAGTGGAGTTTTAGACCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA

[0404] Clone: ​​21E1

[0405] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.102, its encoding nucleic acid is shown in SEQ ID NO.103, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 104 and 105, respectively.

[0406] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGFSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTNYNPSLKS RFS------------FR3------------->CDR3<---FR4--->

[0407] ITRDTSKNQFFLQLNSVTTEDTATYYCAR SYDYWFPY WGQGTLVTVSA

[0408] Nucleic acid sequence

[0409] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTTCTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGCTACAGTGGTAGAACTA ACTACAACCCATCTCTCAAAAGTCGATTCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGATCATATGATTACTGGTTTCCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA

[0410] The amino acid sequence of the light chain VK is shown in SEQ ID NO.106, its encoding nucleic acid is shown in SEQ ID NO.107, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.108, 109 and 110 respectively.

[0411] <----------FR1-------->CDR1<-----FR2----->CDR2<-------DVLMTQTPLSLPVSLGDQASISC RSSQSIVHSNGNTFLE WYLQKPGQSPKLLIY KVSNRFS GVPDRFSG----FR3------------------>CDR3<---FR4--->

[0412] SGSGTDFTLKISRVEAEDLGVYYC FQGSHIPVT FGAGTKLELK

[0413] Nucleic acid sequence

[0414] GATGTTTTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGCATTGTACATAGTAATGGAAACACCTTTTTGGAATGGTACCTGCAGAAACCAGGCCAGTCTCCAAAGCTCCTGATCTACAAAGTT TCCAACCGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGATGGAGGCTGAGGATCTGGGAGTTTATTACTGCTTTCAAGGTTCACATATTCCGGTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0415] Clone: ​​24G17

[0416] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.111, its encoding nucleic acid is shown in SEQ ID NO.112, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.26, 113 and 114, respectively.

[0417] <-------------FR1----------->CDR1<----FR2----->CDR2<--EVQLQESGPGLVKPSQSLSLTCTVTGFSI TSDYAWH WIRQFPGNKLEWMG FINYSGRTNYNPSLTS RIS------------FR3------------->CDR3<---FR4--->

[0418] IIRDTSKNQFFLQLNSVTTEDTATYYCAR SYDFWFGY WGQGTLVTVSA

[0419] Nucleic acid sequence

[0420] GAGGTCCAGCTGCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTTCTCAATCACCAGTGATTATGCCTGGCACTGGATCCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTTCATAAACTACAGTGGTAGAACTA ACTACAACCCATCTCTCACAAGTCGAATCTCTATCATTCGGGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGATCATATGATTTCTGGTTTGGTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA

[0421] The amino acid sequence of the light chain VK is shown in SEQ ID NO.115, its encoding nucleic acid is shown in SEQ ID NO.116, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.117, 118 and 33, respectively.

[0422] <----------FR1-------->CDR1<-----FR2----->CDR2<-------NVLMTQSPLSLPVSLGDQASISC RSSQSIVHSNGNTYLE WYMQKPGQSPKLLIY TVSKRFS GVPDRFSG----FR3------------------>CDR3<---FR4--->

[0423] SGSGTDFTLKISRVEAEDLGVYYC FQGSHVPVT FGAGTKLELK

[0424] Nucleic acid sequence

[0425] aATGTTtTGATGACCcAAAGTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGTATTGTACATAGTAATGGAAACACCTATTTAGAATGGTATATGCAGAAACCAGGCCAGTCTCCAAAGCTCCTGATCTACACAGTT TCCAAACGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGAGTGGAGGCTGAGGATCTGGGAGTTTATTATTGCTTTCAAGGTTCACATGTTCCGGTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0426] Clone: ​​25L15

[0427] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.119, the encoding nucleic acid is shown in SEQ ID NO.120, and the CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.94, 95 and 96, respectively.

[0428] <-------------FR1----------->CDR1<----FR2----->CDR2<--KVKLVESGGDLVKPGGSLKISCAASGFTF SNYAMS WIRQTPDKRLEWVA TVSSGGPYTYYPDSVKG RFT------------FR3------------->CDR3<---FR4--->

[0429] ISRDNAENTLYLQMSSLRSDDTAIYYCAR QIYSDYDVTY WGQGTLVTVSA

[0430] Nucleic acid sequence

[0431] aAAGTGAAGCTGGTGGAGTCTGGGGGAGACTTAGTGAAGCCTGGAGGGTCCCTGAAAATCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAACTATGCCATGTCTTGGATTCGCCAGACTCCGGACAAGAGGCTGGAGTGGGTCGCAACCGTTAGTAGTGGTGGTCCTTACACCTACT ATCCAGACAGTGTGAAGGGGCGATTCACCATCTCCAGAGACAATGCCGAAAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGATGACACGGCCATTTATTACTGTGCAAGGCAGATCTACTCTGATTACGACGTCACTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA

[0432] The amino acid sequence of the light chain VK is shown in SEQ ID NO.121, its encoding nucleic acid is shown in SEQ ID NO.122, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NO.123, 100 and 101 respectively.

[0433] <----------FR1-------->CDR1<-----FR2----->CDR2<-------------QIVLSQSPAILSASPGETVTMTC RVSSSVSYMY WLQQKPGSSPKPWIY ATSNLAS GVPARFSGSGSGTS----FR3---------->CDR3<---FR4--->

[0434] YSLTITRVEAEDAATYYC QQWSFRPWT FGGGTKLEIK

[0435] Nucleic acid sequence

[0436] CAAATTGTTCTTCCCAGTCTCCAGCAATCCTGTCTGCATCTCCAGGGGAGACGGTCACAATGACTTGCAGGGTCAGTTCAAGTGTAAGTTACATGTACTGGCTCCAACAGAAGCCAGGATCCTCCCCCAAACCCTGGATTTATGCCACATCCAACCTG GCTTCTGGAGTCCCTGCTCGCTTCAGTGGCAGTGGGTCTGGGACCTCTTACTCCCTCACAATCACCAGAGTGGAGGCTGAAGATGCTGCCACTTATTACTGTCAGCAGTGGAGTTTTAGACCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA

[0437] Clone: ​​28O14

[0438] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.124, its encoding nucleic acid is shown in SEQ ID NO.125, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.126, 127 and 128 respectively.

[0439] <-------------FR1---------->CDR1<----FR2----->CDR2<--QVQLQQSGTELARPGASVRMSCKASGYTFR SHTMH WVKQRPGQGLEWIG YINPSTGYINYNQKFKD KAT------------FR3------------>CDR3<---FR4--->

[0440] LTADKSSSTAYMQLSSLTSEDSAVYYCAR SNYSYYYGMDY WGQGTSVTVSS

[0441] Nucleic acid sequence

[0442] CAGGTCCAGCTGCAGCAGTCTGGGACTGAACTGGCAAGACCTGGGGCCTCAGTGAGGATGTCCTGCAAGGCTTCTGGCTACACCTTTAGGAGTCACACGATGCACTGGGTAAAACAGAGGCCTGGACAGGGTCTGGAATGGATTGGATACATTAATCCTAGCACTGGTTATATTAATTAC AATCAGAAGTTCAAGGACAAGGCCACATTGACTGCAGACAAATCCTCCAGCACAGCCTACATGCAACTGAGCAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAGAAGTAACTACTCTTATTACTATGGTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0443] The amino acid sequence of the light chain VK is shown in SEQ ID NO.129, its encoding nucleic acid is shown in SEQ ID NO.130, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.131, 132 and 133 respectively.

[0444] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSHKFMSTSVGDRVTITC KASQDVNTAVA WYQQKPGQSPKLLIY SASYRYT GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0445] DFTFTISSLQAEDLAVYYC QQHYNIPLT FGAGTKLELK

[0446] Nucleic acid sequence

[0447] GACATTTGTGATGACCCAGTCTCACAAATTCATGTCCACATCAGTGGGAGACAGGGTCACCATCACCTGCAAGGCCAGTCAGGATGTAAATACTGCTGTAGCCTGGTATCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTACTCGGCATCCTACC GGTACACTGGGGTCCCTGATCGCTTCACTGGCAGTGGATCTGGGACGGATTTCACTTTCACCATCAGCAGTTTACAGGCTGAAGACCTGGCAGTTTTATTACTGTCAGCAACATTATAATATTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0448] The light chain variable region of the above antibody was cloned into a vector containing a human light chain constant region, and the heavy chain variable region was cloned into a vector containing a human heavy chain constant region. The plasmids were co-transfected into CHO-S cells, and the complete IgG antibody was expressed and secreted into the culture medium at 37°C and 5% CO 2 After 7 days of expression at 125 rpm, the supernatant was collected and purified by Protein A chromatography to obtain chimeric IgG, and the concentration of IgG was determined by spectrophotometry. The control antibody 17D20_3521N11 (OMS646) was synthesized according to the document US20120282263.

[0449] According to the method of Example 1, the inhibitory activity of each chimeric antibody on the split C4 reaction was detected, and the OD450nm value was read. After collecting the data, the results were calculated using GraphPad Prism 5 software. The results are shown in FIG. Figure 3 As shown, chimeras 10L3, 3B9, 16A17, 4F18, 25L15, 28O14 and 1B2 had significantly stronger inhibitory activity against MASP-2 than the control antibody in the C4 cleavage activity assay.

[0450] According to the method of Example 1, the C3 convertase activity inhibition test was performed on the selected superior chimeric antibodies, and the results were calculated using GraphPad Prism 5 software after data collection. Figure 4 As shown, in the C3 convertase activity assay, the dominant chimeras 10L3, 3B9, 16A17, 4F18, 25L15, 28O14 and 1B2 showed extremely strong inhibitory activity, which was significantly stronger than the control antibody.

[0451] Affinity evaluation

[0452] 1 μg / mL human MASP-2D4-6 recombinant protein was coated on a 96-well ELISA plate and incubated overnight at 4°C. 2% skim milk was used for blocking at room temperature for 1 hour, and a gradient dilution of MASP-2 antibodies, chimeras 10L3, 3B9, 16A17, 4F18, 25L15, 28O14 and 1B2 (starting concentration of 30 μg / ml, 1:4 gradient dilution) was added, incubated at room temperature for 60 minutes, and Goat anti-human Fc-HRP was added at a dilution ratio of 1:4000, and incubated at room temperature for 60 minutes. TMB was used for color development for 10 minutes, and concentrated sulfuric acid was added to terminate the reaction. OD450nm was measured, and the results were calculated using Graph Pad Prism 5 software.

[0453] The results are as follows Figure 5 As shown, the chimeric antibody binds to human, cynomolgus monkey, rat, and mouse MASP-2 (Table 2).

[0454] Table 2 Binding of MASP-2 antibodies to various proteins

[0455] EC50 (nM) Human MASP-2 Cynomolgus monkey MASP-2 Rat MASP-2 Mouse MASP-2 1B2 0.015 0.014 1.439 0.947 3B9 0.025 0.038 1.624 4.393 4F18 0.004 0.003 0.127 0.287 10L3 0.022 0.016 30.19 30.90 16A17 0.014 0.014 4.296 1.973 25L15 0.012 0.008 0.021 0.023 28O14 0.033 0.043 - -

[0456] Using surface plasmon resonance technology (SPR / Biacore T200), 0.1 μg / ml antibody was captured on the surface of Protein A chip, the capture time was set to 40 seconds, the flow rate was 10 μl / min, the chip was rinsed with buffer until the baseline was stable, and then the gradient diluted antigen was flowed through the chip at a flow rate of 30 μl / min, the binding time was set to 120 seconds, the dissociation time was set to 600 seconds, and the equilibrium dissociation constant (KD) was obtained by fitting with Langmuir 1:1 kinetics mode. The affinity of the candidate antibodies to various MASP-2 antigens is shown in Table 3.

[0457] Table 3 Affinity of MASP-2 antibodies to various genus proteins

[0458] KD (nM) Human MASP-2 Cynomolgus monkey MASP-2 Rat MASP-2 Mouse MASP-2 1B2 0.002 0.078 Weak 632 3B9 0.067 0.051 1043 476 4F18 0.108 24.76 2471 218 10L3 0.070 1.290 Weak 214 16A17 0.008 0.063 Weak 487 25L15 0.120 0.920 167 499 28O14 0.090 0.010 162 767 Control antibody 21.1 8.750 88.5 225

[0459] Example 4: Construction and screening of phage antibody library

[0460] The total RNA of some immune mouse B cells obtained in Example 3 was extracted and reverse transcribed using light chain and heavy chain specific primers to obtain an antibody cDNA library. The variable region fragments of the antibody were specifically amplified using variable region primers and cloned into phage plasmids after restriction digestion to construct an antibody Fab phage display library based on M13 phage with a library capacity of 1×10 10 .

[0461] The library was screened using the traditional phage panning method. MASP-2D4-6 recombinant protein was coated on an immunotube, and approximately 1×10 12 Phages were incubated and panned. After two rounds of panning, single clones were picked and inoculated into 96-well U-shaped plates for culture and induced expression. The culture supernatant was used for ELISA detection of MASP-2 recombinant protein. A total of 494 monoclonal Fab antibodies that can cross-recognize human, cynomolgus monkey, rat and mouse MASP-2 were obtained, of which 34 clones can inhibit C4 deposition catalyzed by MASP-2 ( Figure 6 ).

[0462] After the above Fab clone with inhibitory activity was re-induced to express, the Fab antibody was purified and the inhibitory activity of MASP-2 antibody was tested by C4 cleavage activity assay. Figure 7 As shown, monoclonal antibodies 3H6, 14F11, 16B11, 20C8, 21A3 and 21B1 had the most significant inhibitory activity on MASP-2-catalyzed C4b deposition when the concentration was greater than 0.01 μg / ml.

[0463] Surface plasmon resonance technology (SPR / Biacore T200) was used to capture Anti-Flag antibodies using Protein A chips. 10 μg / ml purified Fab antibodies containing Flag tags were captured on the chip surface. The capture time was set to 40 seconds and the flow rate was 10 μl / min. The chip was rinsed with buffer until the baseline was stable. Then, the gradient diluted MASP-2 antigen was injected as the analyte in sequence to measure the affinity between Fab and antigens of different species. The data were fitted by Biacore T200 Evaluationsoftware 3.1 software in the Langmuir 1:1kinetics mode. Table 4 shows the affinity of monoclonal antibodies 16B11 and 14F11 to MASP-2 antigens of various species.

[0464] Table 4 Affinity of MASP-2 antibody (Fab) and antigen

[0465]

[0466] The variable region sequence of the Fab antibody was obtained by Sanger sequencing as follows:

[0467] Table 5. Variable region sequences of MASP-2 mouse antibodies

[0468]

[0469] Clone: ​​3H6

[0470] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.134, its encoding nucleic acid is shown in SEQ ID NO.135, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.136, 137 and 138 respectively.

[0471] <-------------FR1---------->CDR1<----FR2----->CDR2<--QVQLQQSGTELARPGASVKMSCKASGYTF ATYSIH WVKQRPGQGLEWIG YINPSSGYTNYNQKFRD KAT------------FR3------------>CDR3<---FR4--->

[0472] LTADRSNTAYMQLSSLTSEDSAVYYCAR VYYTFYHAMDY WGQGTSVTVSS

[0473] Nucleic acid sequence

[0474] CAGGTCCAGCTGCAGCAGTCTGGGACTGAACTGGCAAGACCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACCTTTGCTACCTACTCGATACACTGGGTAAAACAGAGGCCTGGGCAGGGTCTGGAATGGATTGGATACATTAATCCTAGCAGTGGTTATACTAATTAC AATCAGAAGTTCAGGGACAAGGCCACATTGACTGCAGACAGATCCTCCAACACAGCCTACATGCAACTGAGCAGCCTGACATCTGAGGACTCTGCAGTCTATTACTGTGCAAGAGTCTACTATACTTTTTACCATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0475] The amino acid sequence of the light chain VK is shown in SEQ ID NO.129, its encoding nucleic acid is shown in SEQ ID NO.130, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.131, 132 and 133 respectively.

[0476] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSHKFMSTSVGDRVTITC KASQDVNTAVA WYQQKPGQSPKLLIY SASYRYT GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0477] DFTFTISSLQAEDLAVYYC QQHYNIPLT FGAGTKLELK

[0478] Nucleic acid sequence

[0479] GACATTTGTGATGACCCAGTCTCACAAATTCATGTCCACATCAGTGGGAGACAGGGTCACCATCACCTGCAAGGCCAGTCAGGATGTAAATACTGCTGTAGCCTGGTATCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTACTCGGCATCCTACC GGTACACTGGGGTCCCTGATCGCTTCACTGGCAGTGGATCTGGGACGGATTTCACTTTCACCATCAGCAGTTTACAGGCTGAAGACCTGGCAGTTTTATTACTGTCAGCAACATTATAATATTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0480] Clone: ​​16B11

[0481] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.139, the encoding nucleic acid thereof is shown in SEQ ID NO.140, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NO.7, 17 and 141, respectively. <-------------FR1----------->CDR1<----FR2----->CDR2<--QMQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG Y ISYSGRTSYNPSLKS RIS-----------FR3--------------->CDR3<---FR4--->

[0482] ITRDTSKNQFFLQLNSVTTEDTATYYCARHYGDY WGQGTTLTVSS

[0483] Nucleic acid sequence

[0484] CAGATGCAGCTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGCTACAGTGGTAGG ACTAGCTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGACACTACGGAGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA

[0485] The amino acid sequence of the light chain VK is shown in SEQ ID NO.142, its encoding nucleic acid is shown in SEQ ID NO.143, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.144, 145 and 146 respectively.

[0486] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKLMSTSVGDRVSVTC KASQNVGTNVA WYQQKPGQSPKALIY SASYRNS GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0487] DFTLTISNVQSEDLAEYFC QQFNTNPLT FGSGTKLEIK

[0488] Nucleic acid sequence

[0489] GACATTTGTGATGACTCAGTCTCAAAAACTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGGTACTAATGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCTAAAGCACTGATTTACTCGGCATCCTACC GGAACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGTTCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTATTTCTGTCAGCAATTTAACACCAATCCTCTCACGTTCGGCTCGGGGACAAAGTTGGAAAATAAAA

[0490] Clone: ​​14F11

[0491] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.147, its encoding nucleic acid is shown in SEQ ID NO.148, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.149, 150 and 151 respectively.

[0492] <-------------FR1---------->CDR1<----FR2----->CDR2<--QVQLQQSGPQLLRPGASVKISCKASGFSF TNHWIH WVKQRPGQGLEWIG MIDPSDSETGLNQKFRD KAT-----------FR3--------------->CDR3<---FR4--->

[0493] LTVDKSSSTAYMHLNSPTSEDSAVYYCAR LGIHDGYYIYAMDF WGQGTSVTVSS

[0494] Nucleic acid sequence

[0495] CAGGTTCAGCTGCAGCAGTCTGGGCCTCAGCTACTTAGGCCTGGGGCTTCAGTGAAGATATCCTGCAAGGCTTCTGGTTTCTCATTCACCAACCACTGGATACACTGGGTGAACAGAGGCCTGGACAAGGTCTTGAGTGGATTGGCATGATTGATCCTTCCGATAGTGAAACTGGCCTCAATC AGAAGTTCAGGGACAAGGCCACATTGACTGTAGACAAGTCCTCCAGCACAGCCTACATGCACCTCAACAGCCCGACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGATTGGGAATCCATGATGGTTACTACATCTATGCTATGGACTTTTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA

[0496] The amino acid sequence of the light chain VK is shown in SEQ ID NO.152, its encoding nucleic acid is shown in SEQ ID NO.153, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.154, 155 and 156 respectively.

[0497] <----------FR1-------->CDR1<-----FR2----->CDR2<------------NIVMTQSPKSMMSVGERVTLSC KASENVGTYVS WYQQKPEQSPKLLIY GASNRYT GVPDRFTGSGSAT----FR3----------->CDR3<---FR4--->

[0498] DFTLTISSVQAEDLADYHC GQSYSYPLT FGAGTKLELK

[0499] Nucleic acid sequence

[0500] AACATTGTAATGACCCAATCTCCCAAATCCATGTCCATGTCAGTAGGAGAGGGTCACCTTGAGCTGCAAGGCCAGTGAGAATGTGGGTACTTATGTATCCTGGTATCAACAGAAACCAGAGCAGTCTCCTAAACTGCTGATATACGGGGCATCCAACC GGTACACTGGGGTCCCCGATCGCTTCACAGGCAGTGGATCTGCAACAGATTTCACTCTGACCATCAGCAGTGTGCAGGCTGAAGACCTTGCAGATTATCACTGTGGACAGAGTTACAGCTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0501] Clone: ​​20C8

[0502] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.157, its encoding nucleic acid is shown in SEQ ID NO.158, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 9 respectively.

[0503] <-------------FR1---------->CDR1<----FR2----->CDR2<--EVQLQESGPGLVKPSQSLSLTCSVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPSLKS RIS-----------FR3--------------->CDR3<---FR4--->

[0504] ITRDTSKNQFFLQLISVTTEDTATYYCAR YWGDY WGQGTTLTVSS

[0505] Nucleic acid sequence

[0506] GAGGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCTCTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTACAGTGGTAGA ACTAGCTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGATTTCTGTGACTACTGAGGACACAGCCACATATTATTGTGCAAGATATTGGGGCGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA

[0507] The amino acid sequence of the light chain VK is shown in SEQ ID NO.159, its encoding nucleic acid is shown in SEQ ID NO.160, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.161, 162 and 14, respectively.

[0508] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFISTSIGDRVSVTC KASQDVGGHVA WFQQRPGQSPKPLIY SATFRHS GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0509] DFTLTISNLQSEDLAEYVC LQYNSNPLT FGPGTKLELK

[0510] Nucleic acid sequence

[0511] GACATTGTGATGACCCAGTCTCAAAAATTCATCTCCACATCAATAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGGATGTGGGTGGACATGTGGCCTGGTTTCAACAGAGACCCGGGCAATCTCCTAAACCTCTGATTTATTCGGCAACCTTCC GGCACAGTGGAGTCCCTGATCGCTTCACGGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGTAATCTACAGTCTGAGGACTTGGCAGAATATGTCTGTCTGCAATATAACAGTAATCCACTCACGTTCGGTCCTGGGACCAAGCTGGAGCTGAAA

[0512] Clone: ​​21A3

[0513] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.163, its encoding nucleic acid is shown in SEQ ID NO.164, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 165 and 9, respectively.

[0514] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPYLKS RIS------------FR3------------->CDR3<---FR4--->

[0515] ITRDTSKNQFFLQLNSVTTEDTATYYCAR YWGDY WGQGTTLTVSS

[0516] Nucleic acid sequence

[0517] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGCTACAGTGGTCGC ACTAGCTACAACCCATATCTCAAAAGTCGAATTTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTACTGAGGACACAGCCACATATTACTGTGCAAGATATTGGGGCGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA

[0518] The amino acid sequence of the light chain VK is shown in SEQ ID NO.166, its encoding nucleic acid is shown in SEQ ID NO.167, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.144, 168 and 14, respectively.

[0519] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFMSTSVGDRVSVTC KASQNVGTNVA WYKQKPGQSPKALIY SASYRHS GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0520] DFTLTISNVQSEDLAEYFC LQYNSNPLT FGAGTKLELK

[0521] Nucleic acid sequence

[0522] GACATTTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGGTACTAATGTAGCCTGGTATAAACAGAAACCAGGGCAATCTCCTAAAGCACTGATTTACTCGGCATCCTACC GGCACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTATTTCTGTCTGCAATATAACAGCAATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0523] Clone: ​​21B1

[0524] The amino acid sequence of the heavy chain VH is shown in SEQ ID NO.169, its encoding nucleic acid is shown in SEQ ID NO.170, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.7, 17 and 9 respectively.

[0525] <-------------FR1----------->CDR1<----FR2----->CDR2<--DVQLQESGPGLVKPSQSLSLTCTVTGYSI TSDYAWN WIRQFPGNKLEWMG YISYSGRTSYNPSLKS RIS------------FR3------------->CDR3<---FR4--->

[0526] ITRDTSKNQFFLQLNSVTAEDTATYYCAR YWGDY WGQGTTLTVSS

[0527] Nucleic acid sequence

[0528] GATGTGCAGCTTCAGGAGTCGGGACCTGGCCTGGTGAAACCTTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTATGCCTGGAACTGGATCCGGCAGTTTCCAGGAAACAAACTGGAGTGGATGGGCTACATAAGCTACAGTGGTCGC ACTAGCTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGACTGCTGAGGACACAGCCACATATTACTGTGCAAGATATTGGGGCGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA

[0529] The amino acid sequence of the light chain VK is shown in SEQ ID NO.171, its encoding nucleic acid is shown in SEQ ID NO.172, and its CDR1, CDR2 and CDR3 are shown in SEQ ID NOs.144, 173 and 174 respectively.

[0530] <----------FR1-------->CDR1<-----FR2----->CDR2<------------DIVMTQSQKFMSTSVGDRVSVTC KASQNVGTNVA WFQQKPGQSPKALIY STSYRSS GVPDRFTGSGSGT----FR3----------->CDR3<---FR4--->

[0531] DFTLTISNVQSEDLAEYFC QQFNSNPLT FGGGTKLELK

[0532] Nucleic acid sequence

[0533] GACATTTGTGATGACACAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGGTACTAATGTAGCCTGGTTTCAACAGAAACCAGGGCAATCTCCTAAAGCACTGATTTACTCGACATCCTACC GGTCCAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTACTTCTGTCAGCAATTTAACAGCAATCCTCTCACGTTCGGAGGGGGGACCAAGCTGGAGCTGAAA

[0534] Example 5. Humanization of MASP-2 Antibodies

[0535] Mouse antibodies 28O14, 3H6 and 16B11 with high affinity and strong activity were selected for variable region sequence humanization. First, the glycosylation site NYS of the heavy chain CDR3 of 28O14 was cloned and site-directed mutagenesis was performed. The mutant was expressed as Fab in E. coli prokaryotes. Affinity detection showed that after the glycosylation site of the heavy chain CDR3 of 28O14 was removed, the SA mutant (Ala102) in which the 102nd position was mutated from serine to alanine did not affect the MASP-2 binding activity (such as Figure 8 shown).

[0536] The mouse antibody sequence was compared with the human antibody germline sequence to find the human germline light chain gene with good homology, the same key amino acid sequence that maintains the core of the antibody structure, and the high frequency in the human body, and then the mouse antibody CDR was transplanted; homology modeling was performed by computer to analyze the CDR region and its surrounding framework amino acid sequence to avoid the concentrated distribution of molecular surface charge or hydrophobic regions. The humanized design obtained 3 heavy chain and 6 light chain 16B11 variants, 4 heavy chain and 1 light chain 3H6 variants, 4 heavy chain and 1 light chain 28O14 variants, and 3 heavy chain and 6 light chain 16B11 variants.

[0537] The humanized sequences are as follows:

[0538] Table 6. Heavy chain variable region sequences of MASP-2 humanized antibodies

[0539]

[0540] Table 7. Light chain variable region sequences of MASP-2 humanized antibodies

[0541]

[0542] h16B11:

[0543] h16B11 H1

[0544] The amino acid sequence of h16B11 H1 is shown in SEQ ID NO.175, the encoding nucleic acid thereof is shown in SEQ ID NO.176, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.7, 17 and 141, respectively.

[0545] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QMQLQESGPGLVKPSETLSLTCTVSGYSI TSDYAWN WIRQPPGKGLEWIG YISYSGRTSYNPSLKS RVTISRD----------FR3---------->CDR3<----FR4--->

[0546] TSKNQFSLKLSSVTAADTAVYYCAR HYGDY WGQGTLVTVSS

[0547] Nucleic acid sequence

[0548] CAGATGCAGCTGCAAGAGTCTGGACCTGGACTGGTCAAGCCCAGCGAGACACTGAGCCTGACCTGTACCGTGTCCGGCTACAGCATCACAAGCGACTACGCCTGGAACTGGATCAGACAGCCTCCTGGCAAAGGCCTCGAGTGGATCGGCTACATCAGCTACAGCGGCAGA ACCAGCTACAACCCCAGCCTGAAGTCCAGAGTGACCATCAGCAGAGACACCAGCAAGAACCAGTTCTCCCTGAAGCTGAGCAGCGTGACAGCCGCCGATACAGCCGTGTACTACTGCGCTAGACACTACGGCGATTACTGGGGCCAGGGAACCCTGGTCACAGTGTCATCT

[0549] h16B11 H2

[0550] The amino acid sequence of h16B11 H2 is shown in SEQ ID NO.177, the encoding nucleic acid thereof is shown in SEQ ID NO.178, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.7, 17 and 141, respectively.

[0551] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QLQLQESGPGLVKPSETLSLTCTVSGYSI TSDYAWN WIRQPPGKGLEWIG YISYSGRTSYNPSLKS RVTISRD----------FR3------------->CDR3<----FR4--->

[0552] TSKNQFSLKLSSVTAADTAVYYCAR HYGDY WGQGTLVTVSS

[0553] Nucleic acid sequence

[0554] CAGCTCCAGCTGCAAGAGTCTGGACCTGGACTGGTCAAGCCCAGCGAGACACTGAGCCTGACCTGTACCGTGTCCGGCTACAGCATCACAAGCGACTACGCCTGGAACTGGATCAGACAGCCTCCTGGCAAAGGCCTCGAGTGGATCGGCTACATCAGCTACAGCGGCAGA ACCAGCTACAACCCCAGCCTGAAGTCCAGAGTGACCATCAGCAGAGACACCAGCAAGAACCAGTTCTCCCTGAAGCTGAGCAGCGTGACAGCCGCCGATACAGCCGTGTACTACTGCGCTAGACACTACGGCGATTACTGGGGCCAGGGAACCCTGGTCACAGTGTCATCT

[0555] h16B11 H3

[0556] The amino acid sequence of h16B11 H3 is shown in SEQ ID NO.179, the encoding nucleic acid thereof is shown in SEQ ID NO.180, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.7, 17 and 141, respectively.

[0557] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QLQLQESGPGLVKPSETLSLTCTVSGYSI TSDYAWN WIRQPPGKGLEWIG YISYSGRTSYNPSLKS RVTISVD----------FR3------------->CDR3<----FR4--->TSKNQFSLKLSSVTAADTAVYYCAR HYGDY WGQGTLVTVSS

[0558] Nucleic acid sequence

[0559] CAGCTCCAGCTGCAAGAGTCTGGACCTGGACTGGTCAAGCCCAGCGAGACACTGAGCCTGACCTGTACCGTGTCCGGCTACAGCATCACAAGCGACTACGCCTGGAACTGGATCAGACAGCCTCCTGGCAAAGGCCTCGAGTGGATCGGCTACATCAGCTACAGCGGCAGA ACCAGCTACAACCCCAGCCTGAAGTCCAGAGTGACCATCAGCgtAGACACCAGCAAGAACCAGTTCTCCCTGAAGCTGAGCAGCGTGACAGCCGCCGATACAGCCGTGTACTACTGCGCTAGACACTACGGCGATTACTGGGGCCAGGGAACCCTGGTCACAGTGTCATCT

[0560] h3H6:

[0561] h3H6 H1

[0562] The amino acid sequence of h3H6 H1 is shown in SEQ ID NO.181, the encoding nucleic acid thereof is shown in SEQ ID NO.182, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.136, 183 and 138, respectively.

[0563] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF ATYSIH WVRQAPGQGLEWMG YINPSSGYTNYNQKFQGRVTLTAD--------FR3-------------->CDR3<----FR4--->KSTSTAYMELSSLRSEDTAVYYCAR VYYTFYHAMDY WGQGTTVTVSS

[0564] Nucleic acid sequence

[0565] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCGCCACCTACAGCATCCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCTCTGGCTACACCAACTAC AACCAGAAATTCCAGGGCCGCGTGACCcTCACAGCCGACaAGTCTACAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAGTGTACTACACATTCTACCACGCCATGGACTACTGGGGCCAGGGCACAACAGTGACCGTTAGCTCT

[0566] h3H6 H2

[0567] The amino acid sequence of h3H6 H2 is shown in SEQ ID NO.184, the encoding nucleic acid thereof is shown in SEQ ID NO.185, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.136, 183 and 138, respectively.

[0568] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF ATYSIH WVRQAPGQGLEWMG YINPSSGYTNYNQKFQG RVTLTAD--------FR3-------------->CDR3<----FR4--->

[0569] ESTSTAYMELSSLRSEDTAVYYCAR VYYTFYHAMDY WGQGTTVTVSS nucleic acid sequence

[0570] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCGCCACCTACAGCATCCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCTCTGGCTACACCAACTAC AACCAGAAATTCCAGGGCCGCGTGACCcTCACAGCCGACGAGTCtacAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAGTGTACTACACATTCTACCACGCCATGGACTACTGGGGCCAGGGCACAACAGTGACCGTTAGCTCT

[0571] h3H6 H3

[0572] The amino acid sequence of h3H6 H3 is shown in SEQ ID NO.186, the encoding nucleic acid thereof is shown in SEQ ID NO.187, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.136, 183 and 138, respectively.

[0573] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF ATYSIH WVRQAPGQGLEWMG YINPSSGYTNYNQKFQG RVTITAD--------FR3-------------->CDR3<----FR4--->

[0574] KSTSTAYMELSSLRSEDTAVYYCAR VYYTFYHAMDY WGQGTTVTVSS

[0575] Nucleic acid sequence

[0576] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCGCCACCTACAGCATCCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCTCTGGCTACACCAACTAC AACCAGAAATTCCAGGGCCGCGTGACACAGCCGACaAGTCTACAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAGTGTACTACACATTCTACCACGCCATGGACTACTGGGGCCAGGGCACAACAGTGACCGTTAGCTCT

[0577] h3H6 H4

[0578] The amino acid sequence of h3H6 H4 is shown in SEQ ID NO.188, the encoding nucleic acid thereof is shown in SEQ ID NO.189, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.136, 183 and 138, respectively.

[0579] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF ATYSIH WVRQAPGQGLEWMG YINPSSGYTNYNQKFQG RVTITAD--------FR3-------------->CDR3<----FR4--->

[0580] ESTSTAYMELSSLRSEDTAVYYCAR VYYTFYHAMDY WGQGTTVTVSS nucleic acid sequence

[0581] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCGCCACCTACAGCATCCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCTCTGGCTACACCAACTAC AACCAGAAATTCCAGGGCCGCGTGACACAGCCGACGAGTCTACAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAGTGTACTACACATTCTACCACGCCATGGACTACTGGGGCCAGGGCACAACAGTGACCGTTAGCTCT

[0582] h28O14:

[0583] h28O14 H1

[0584] The amino acid sequence of h28O14 H1 is shown in SEQ ID NO.190, the encoding nucleic acid thereof is shown in SEQ ID NO.191, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.126, 192 and 128, respectively.

[0585] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF RSHTMH WVRQAPGQGLEWMG YINPSTGYINYNQKFQG RVTLTAD--------FR3-------------->CDR3<----FR4--->KSTSTAYMELSSLRSEDTAVYYCAR SNYAYYYGMDY WGQGTTVTVSS

[0586] Nucleic acid sequence

[0587] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCAGATCCCACACCATGCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCACCGGCTATATCAACTAC AACCAGAAATTCCAGGGCCGCGTGACCcTCACAGCCGACaAGTCTACAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAAGCAACTACGCCTACTACTACGGCATGGACTACTGGGGCCAGGGCACCACAGTGACAGTCTCTTCT

[0588] h28O14 H2

[0589] The amino acid sequence of h28O14 H2 is shown in SEQ ID NO.193, the encoding nucleic acid thereof is shown in SEQ ID NO.194, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.126, 192 and 128, respectively.

[0590] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF RSHTMH WVRQAPGQGLEWMG YINPSTGYINYNQKFQG RVTLTAD--------FR3-------------->CDR3<----FR4--->ESTSTAYMELSSLRSEDTAVYYCAR SNYAYYYGMDY WGQGTTVTVSS

[0591] Nucleic acid sequence

[0592] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCAGATCCCACACCATGCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCACCGGCTATATCAACTAC AACCAGAAATTCCAGGGCCGCGTGACCcTCACAGCCGACGAGTCtacAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAAGCAACTACGCCTACTACTACGGCATGGACTACTGGGGCCAGGGCACCACAGTGACAGTCTCTTCT

[0593] h28O14 H3

[0594] The amino acid sequence of h28O14 H3 is shown in SEQ ID NO.195, the encoding nucleic acid thereof is shown in SEQ ID NO.196, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.126, 192 and 128, respectively.

[0595] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF RSHTMH WVRQAPGQGLEWMG YINPSTGYINYNQKFQG RVTITAD--------FR3-------------->CDR3<----FR4--->

[0596] KSTSTAYMELSSLRSEDTAVYYCAR SNYAYYYGMDY WGQGTTVTVSS nucleic acid sequence

[0597] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCAGATCCCACACCATGCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCACCGGCTATATCAACTAC AACCAGAAATTCCAGGGCCGCTGACCATCACAGCCGACaAGTCTACAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAAGCAACTACGCCTACTACTACGGCATGGACTACTGGGGCCAGGGCACCACAGTGACAGTCTCTTCT

[0598] h28O14 H4

[0599] The amino acid sequence of h28O14 H4 is shown in SEQ ID NO.197, the encoding nucleic acid thereof is shown in SEQ ID NO.198, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.126, 192 and 128, respectively.

[0600] <-------------FR1------------->CDR1<-----FR2----->CDR2<-------QVQLVQSGAEVKKPGSSVKVSCKASGYTF RSHTMH WVRQAPGQGLEWMG YINPSTGYINYNQKFQG RVTITAD--------FR3-------------->CDR3<----FR4--->ESTSTAYMELSSLRSEDTAVYYCAR SNYAYYYGMDY WGQGTTVTVSS

[0601] Nucleic acid sequence

[0602] CAGGTTCAGCTGGTTCAGTCTGGCGCCGAAGTGAAGAAACCTGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTAGCGGCTACACCTTCAGATCCCACACCATGCACTGGGTCCGACAGGCTCCAGGACAGGGACTTGAGTGGATGGGCTACATCAACCCCAGCACCGGCTATATCAACTAC AACCAGAAATTCCAGGGCCGCGTGACACAGCCGACGAGTCTACAAGCACCGCCTACATGGAACTGAGCAGCCTGAGAAGCGAGGACACCGCCGTGTACTACTGCGCCAGAAGCAACTACGCCTACTACTACGGCATGGACTACTGGGGCCAGGGCACCACAGTGACAGTCTCTTCT

[0603] h16B11:

[0604] h16B11 K1

[0605] The amino acid sequence of h16B11 K1 is shown in SEQ ID NO.199, the encoding nucleic acid thereof is shown in SEQ ID NO.200, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.201, 202 and 146, respectively.

[0606] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------DIQMTQSPSSSLSASVGDRVTITC RASQNVGTNVA WYQQKPGKAPKALIY SASYRES GVPSRFSGSGSGTDFTL--FR3---------->CDR3<---FR4--->TISSLQPEDFATYFC QQFNTNPLT FGGGTKVEIK

[0607] Nucleic acid sequence

[0608] GACATCCAGATGACACAGAGCCCTAGCAGCCTGTCTGCCAGCGTGGGAGACAGAGTGACCATCACCTGTAGAGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGgcGCTGATCTACAGCGCCAGCTAcA GAgAGAGCGGCGTGCCATCTAGATTCAGCGGCTCTGGCTCTGGCACCGACTTCACCCTGACAATCTCTAGCCTGCAGCCTGAGGACTTCGCTACCTACTTCTGCCAGCAGTTCAACACAAACCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0609] h16B11 K2

[0610] The amino acid sequence of h16B11 K2 is shown in SEQ ID NO.203, the encoding nucleic acid thereof is shown in SEQ ID NO.204, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.201, 202 and 146, respectively.

[0611] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------DIQMTQSPSSSLSASVGDRVTITC RASQNVGTNVA WYQQKPGKAPKLLIY SASYRES GVPSRFSGSGSGTDFTL--FR3---------->CDR3<---FR4--->TISSLQPEDFATYFC QQFNTNPLT FGGGTKVEIK

[0612] Nucleic acid sequence

[0613] GACATCCAGATGACACAGAGCCCTAGCAGCCTGTCTGCCAGCGTGGGAGACAGAGTGACCATCACCTGTAGAGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGCTGCTGATCTACAGCGCCAGCTACA GAgAGAGCGGCGTGCCATCTAGATTCAGCGGCTCTGGCTCTGGCACCGACTTCACCCTGACAATCTCTAGCCTGCAGCCTGAGGACTTCGCTACCTACTTCTGCCAGCAGTTCAACACAAACCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0614] h16B11 K3

[0615] The amino acid sequence of h16B11 K3 is shown in SEQ ID NO.205, the encoding nucleic acid thereof is shown in SEQ ID NO.206, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.201, 207 and 146, respectively.

[0616] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------DIQMTQSPSSSLSASVGDRVTITC RASQNVGTNVA WYQQKPGKAPKALIY SASYRQS GVPSRFSGSGSGTDFTL--FR3---------->CDR3<---FR4--->TISSLQPEDFATYFC QQFNTNPLT FGGGTKVEIK

[0617] Nucleic acid sequence

[0618] GACATCCAGATGACACAGAGCCCTAGCAGCCTGTCTGCCAGCGTGGGAGACAGAGTGACCATCACCTGTAGAGCCAGCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTGGCAAGGCTCCCAAGgcGCTGATCTACAGCGCCAGCTACA GACAGAGCGGCGTGCCATCTAGATTCAGCGGCTCTGCCTCTGGCACCGACTTCACCCTGACAATCTCTAGCCTGCAGCCTGAGGACTTCGCTACCTACTTCTGCCAGCAGTTCAACACAAACCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0619] h16B11 K4

[0620] The amino acid sequence of h16B11 K4 is shown in SEQ ID NO.208, the encoding nucleic acid thereof is shown in SEQ ID NO.209, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.201, 202 and 146, respectively.

[0621] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------EIVMTQSPATLSVSPGERATLSC RASQNVGTNVA WYQQKPGQAPRALIY SASYRES GIPARFSGSGSGTEFTL--FR3---------->CDR3<---FR4--->TISSLQSEDFAVYFC QQFNTNPLT FGGGTKVEIK

[0622] Nucleic acid sequence

[0623] GAGATCGTGaTGACACAGAGCCCCGCCACACTGAGTGTGTCTCCAGGCGAAAGAGCTACCCTGAGCTGTAGAGCCTTCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTGGACAGGCTCCCAGAgcGCTGATCTACAGCGCCAGCTACA GagagTCTGGCATCCCTGCTAGATTCAGCGGCTCTGGCTCTGGCACCGAGTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGGACTTCGCCGTGTACTTCTGCCAGCAGTTCAACACAAACCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0624] h16B11 K5

[0625] The amino acid sequence of h16B11 K5 is shown in SEQ ID NO.210, the encoding nucleic acid thereof is shown in SEQ ID NO.211, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.201, 202 and 146, respectively.

[0626] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------EIVMTQSPATLSVSPGERATLSC RASQNVGTNVA WYQQKPGQAPRLLIY SASYRES GIPARFSGSGSGTEFTL--FR3---------->CDR3<---FR4--->TISSLQSEDFAVYFC QQFNTNPLT FGGGTKVEIK

[0627] Nucleic acid sequence

[0628] GAGATCGTGaTGACACAGAGCCCCGCCACACTGAGTGTGTCTCCAGGCGAAAGAGCTACCCTGAGCTGTAGAGCCTTCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTGGACAGGCTCCCAGACTGCTGATCTACAGCGCCAGCTACA GagagTCTGGCATCCCTGCTAGATTCAGCGGCTCTGGCTCTGGCACCGAGTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGGACTTCGCCGTGTACTTCTGCCAGCAGTTCAACACAAACCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0629] h16B11 K6

[0630] The amino acid sequence of h16B11 K6 is shown in SEQ ID NO.212, the encoding nucleic acid thereof is shown in SEQ ID NO.213, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.201, 207 and 146, respectively.

[0631] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------EIVMTQSPATLSVSPGERATLSC RASQNVGTNVA WYQQKPGQAPRALIY SASYRQS GIPARFSGSGSGTEFTL--FR3---------->CDR3<---FR4--->

[0632] TISSLQSEDFAVYFC QQFNTNPLT FGGGTKVEIK nucleic acid sequence

[0633] GAGATCGTGaTGACACAGAGCCCCGCCACACTGAGTGTGTCTCCAGGCGAAAGAGCTACCCTGAGCTGTAGAGCCTTCCAGAACGTGGGCACAAACGTGGCCTGGTATCAGCAGAAGCCTGGACAGGCTCCCAGAgcGCTGATCTACAGCGCCAGCTACA GAcagTCTGGCATCCCTGCTAGATTCAGCGGCTCTGCCTCTGGCACCGAGTTCACCCTGACAATCAGCAGCCTGCAGAGCGAGGACTTCGCCGTGTACTTCTGCCAGCAGTTCAACACAAACCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0634] h28O14:

[0635] h28O14 K1

[0636] The amino acid sequence of h28O14 K1 is shown in SEQ ID NO.214, the encoding nucleic acid thereof is shown in SEQ ID NO.215, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.216, 132 and 133, respectively.

[0637] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------DIQMTQSPSSSLSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLLIY SASYRYT GVPSRFSGSGSGTDFTL--FR3---------->CDR3<---FR4--->TISSLQPEDFATYYC QQHYNIPLT FGGGTKVEIK

[0638] Nucleic acid sequence

[0639] GACATCCAGATGACACAGAGCCCTAGCAGCCTGTCTGCCAGCGTGGGAGACAGAGTGACCATCACCTGTAGAGCCAGCCAGGACGTGAACACAGCCGTGGCTTGGTATCAGCAGAAGCCTGGCAAGGCCCCTAAGCTGCTGATCTACAGCGCCAGCTACA GATACACCGGCGTGCCCTCTAGATTCAGCGGCTCTGCCTCTGGCACCGACTTCACCCTGACAATCTCTAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACAACATCCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0640] h3H6 K:

[0641] h3H6 K1

[0642] The amino acid sequence of h3H6 K1 is shown in SEQ ID NO.214, the encoding nucleic acid thereof is shown in SEQ ID NO.215, and the CDR1, CDR2 and CDR3 thereof are shown in SEQ ID NOs.216, 132 and 133, respectively.

[0643] <----------FR1--------->CDR1<-----FR2------>CDR2<----------------DIQMTQSPSSSLSASVGDRVTITC RASQDVNTAVA WYQQKPGKAPKLLIY SASYRYT GVPSRFSGSGSGTDFTL--FR3---------->CDR3<---FR4--->TISSLQPEDFATYYC QQHYNIPLT FGGGTKVEIK

[0644] Nucleic acid sequence

[0645] GACATCCAGATGACACAGAGCCCTAGCAGCCTGTCTGCCAGCGTGGGAGACAGAGTGACCATCACCTGTAGAGCCAGCCAGGACGTGAACACAGCCGTGGCTTGGTATCAGCAGAAGCCTGGCAAGGCCCCTAAGCTGCTGATCTACAGCGCCAGCTACA GATACACCGGCGTGCCCTCTAGATTCAGCGGCTCTGCCTCTGGCACCGACTTCACCCTGACAATCTCTAGCCTGCAGCCTGAGGACTTCGCCACCTACTACTGCCAGCAGCACTACAACATCCCTCTGACCTTCGGCGGAGGCACCAAGGTGGAAATCAAG

[0646] After the full sequences of the light and heavy chains were synthesized, they were cloned into a eukaryotic expression vector containing the antibody kappa chain constant region Ckappa or the human IgG4 constant region CH1-CH3. The light and heavy chain plasmids were combined and paired, and then transfected into CHO-S cells. The cells were expressed at 37°C for 5-6 days, and the culture supernatant was collected and purified using a Protein A column.

[0647] Example 6. MASP-2 Inhibitory Activity of Humanized Antibodies

[0648] C4 cleavage activity assay

[0649] According to the C4 cleavage assay in Example 1, different concentrations of MASP-2 antibody (starting concentration of 30 μg / ml, 5-fold gradient dilution, a total of 9 gradient dilutions) were added to the reaction wells at 50 μl per well and allowed to react at 4°C for 30 minutes. 50 μl of 2 μg / ml C4 purified protein (CompTech) was added and allowed to react at 37°C for 90 minutes. C4c antibody was added to detect C4b deposition. Fig. 9A , 9B As shown in Table 8, the humanized antibodies can significantly inhibit the activity of MASP-2.

[0650] Table 8. Inhibitory activity of MASP-2 humanized antibodies on C4b deposition

[0651]

[0652]

[0653] C3 convertase activity assay

[0654] According to the C3 convertase activity assay method in Example 1, different concentrations of MASP-2 antibody (starting concentration of 30 μg / ml, 5-fold gradient dilution, a total of 9 gradient dilutions) were added to the reaction wells at 50 μl per well and reacted at 37°C for 90 minutes. 1:500 C3b antibody was added to detect C3b deposition. Fig. 10A , 10B As can be seen from 10C and Table 9, each humanized antibody can inhibit the deposition of C3b.

[0655] Table 9. Inhibitory activity of MASP-2 humanized antibodies on C3b deposition

[0656] Humanized Antibodies Heavy chain variable region Light chain variable region IC50(nM) h3H6 H1K1 h3H6 H1 h3H6 K1 0.030 h3H6 H2K1 h3H6 H2 h3H6 K1 0.151 h3H6 H3K1 h3H6 H3 h3H6 K1 0.034 h3H6 H4K1 h3H6 H4 h3H6 K1 0.151 h28O14H1K1 h28O14 H1 h28O14 K1 0.146 h28O14H2K1 h28O14 H2 h28O14 K1 0.483 h28O14H3K1 h28O14 H3 h28O14 K1 0.161 h28O14H4K1 h28O14 H4 h28O14 K1 0.506 h16B11H1K1 h16B11 H1 h16B11 K1 0.014 h16B11H1K2 h16B11 H1 h16B11 K2 0.040 h16B11H1K3 h16B11 H1 h16B11 K3 0.023 h16B11H2K3 h16B11 H2 h16B11 K3 0.018 h16B11H2K1 h16B11 H2 h16B11 K1 0.017 h16B11H2K2 h16B11 H2 h16B11 K2 0.028 h16B11H3K1 h16B11 H3 h16B11 K1 0.060 h16B11H3K2 h16B11 H3 h16B11 K2 0.399 h16B11H3K3 h16B11 H3 h16B11 K3 0.112 h16B11H1K4 h16B11 H1 h16B11 K4 0.015 h16B11H1K5 h16B11 H1 h16B11 K5 0.024 h16B11H1K6 h16B11 H1 h16B11 K6 0.018 h16B11H2K4 h16B11 H2 h16B11 K4 0.015 h16B11H2K5 h16B11 H2 h16B11 K5 0.032 h16B11H2K6 h16B11 H2 h16B11 K6 0.018 h16B11H3K4 h16B11 H3 h16B11 K4 0.052 h16B11H3K5 h16B11 H3 h16B11 K5 0.440 h16B11H3K6 h16B11 H3 h16B11 K6 0.071 Control Antibodies 3.692

[0657] Example 7. Affinity of MASP-2 Humanized Antibodies

[0658] (1) ELISA to detect the affinity of humanized MASP-2 antibodies to various antigens

[0659] 1 μg / mL human MASP-2D4-6 recombinant protein was coated on a 96-well ELISA plate and incubated overnight at 4°C. 2% skim milk was used for blocking at room temperature for 1 hour, and a gradient dilution of MASP-2 antibody (starting concentration was 30 μg / ml, 1:4 gradient dilution) was added, incubated at room temperature for 60 minutes, and a 1:4000 dilution of goat anti-human Fc-HRP was added, and incubated at room temperature for 60 minutes. TMB was used for color development for 10 minutes, and concentrated sulfuric acid was added to terminate the reaction. OD450nm was measured, and the results were calculated using Graph Pad Prism 5 software.

[0660] The results are as follows Fig.11 As shown, the MASP-2 humanized antibody binds to human, cynomolgus monkey, rat, and mouse MASP-2 with comparable affinity.

[0661] (2) BIAcore detection of affinity of humanized antibodies for human and mouse MASP-2

[0662] 0.1 μg / ml antibody was captured on the surface of Protein A chip, the capture time was set to 40 seconds, the flow rate was 10 μl / min, the chip was rinsed with buffer until the baseline was stable, and then the gradient diluted antigen was flowed through the chip at a flow rate of 30 μl / min, the binding time was set to 120 seconds, the dissociation time was set to 600 seconds, and the equilibrium dissociation constant (KD) was obtained by fitting. The affinity determination values ​​are shown in Table 10.

[0663] Table 10. Affinity of MASP-2 Antibodies to MASP-2

[0664]

[0665] Example 7: Shiga toxin 2-induced hemolytic uremic syndrome model in mice

[0666] Using the cDNA plasmid of STX2 (Sino Biological, catalog number: EG40019-G) as a template, the STX2A (Arg23-Lys319) and STX2B (Ala20-Asp89) gene fragments of E. coli O157:H7 were PCR amplified and cloned into the prokaryotic expression plasmid, in which the twin strep tag was fused to the N-terminus of the STX2A protein for purification, and the enterokinase cleavage site DDDDK was added between the two. The constructed STX2 expression plasmid was transformed into TG1 Escherichia coli competent cells, and a single clone was picked out and placed in bacterial culture medium. IPTG was added for induction culture overnight, and the bacteria were collected. PPB and 5 mM MgSO4 solution were added successively to lyse the bacterial cell wall. The protein expression solution in the periplasmic cavity was collected by centrifugation, and the Shiga toxin STX2 protein was purified by Strep-TactinX (Sino HaiGene, Catalog No.: C0402). EK enterokinase was added to the protein collection solution, incubated at 25°C overnight, and the twin strep tag was removed.

[0667] C57 BL / 6 female mice weighing about 18g were selected and divided into 4 dose groups of MASP-2 antibody 0mg / kg, 3 / mg / kg, 10mg / kg and 30mg / kg, with 7 mice in each group. Anti-MASP-2 antibody h16B11 H2K4 (100μl) was injected into the tail vein 2 hours in advance, and LPS (5μg / mouse, L6136-Sigma) and STX2 (10ng / mouse) were injected intraperitoneally afterwards. The survival rate of mice was monitored starting 60 hours after administration, and they were killed when they reached the coma stage of HUS pathology. The results are shown in Fig.12 As shown, anti-MASP-2 antibodies can significantly prolong the survival time of mice in a dose-dependent manner. SEQUENCE LISTING <110> Conoya Biopharmaceutical Technology (Chengdu) Co., Ltd. <120> Development and application of a complement inhibitor <130> MTI20213 <160> 216 <170> PatentIn version 3.5 <210> 1 <211> 397 <212> PRT <213> Homo sapiens <400> 1 His His His His His His Ala Ser Gln Pro Cys Pro Tyr Pro Met Ala 1 5 10 15 Pro Pro Asn Gly His Val Ser Pro Val Gln Ala Lys Tyr Ile Leu Lys 20 25 30 Asp Ser Phe Ser Ile Phe Cys Glu Thr Gly Tyr Glu Leu Leu Gln Gly 35 40 45 His Leu Pro Leu Lys Ser Phe Thr Ala Val Cys Gln Lys Asp Gly Ser 50 55 60 Trp Asp Arg Pro Met Pro Ala Cys Ser Ile Val Asp Cys Gly Pro Pro 65 70 75 80 Asp Asp Leu Pro Ser Gly Arg Val Glu Tyr Ile Thr Gly Pro Gly Val 85 90 95 Thr Thr Tyr Lys Ala Val Ile Gln Tyr Ser Cys Glu Glu Thr Phe Tyr 100 105 110 Thr Met Lys Val Asn Asp Gly Lys Tyr Val Cys Glu Ala Asp Gly Phe 115 120 125 Trp Thr Ser Ser Lys Gly Glu Lys Ser Leu Pro Val Cys Glu Pro Val 130 135 140 Cys Gly Leu Ser Ala Arg Thr Thr Gly Gly Arg Ile Tyr Gly Gly Gln 145 150 155 160 Lys Ala Lys Pro Gly Asp Phe Pro Trp Gln Val Leu Ile Leu Gly Gly 165 170 175 Thr Thr Ala Ala Gly Ala Leu Leu Tyr Asp Asn Trp Val Leu Thr Ala 180 185 190 Ala His Ala Val Tyr Glu Gln Lys His Asp Ala Ser Ala Leu Asp Ile 195 200 205 Arg Met Gly Thr Leu Lys Arg Leu Ser Pro His Tyr Thr Gln Ala Trp 210 215 220 Ser Glu Ala Val Phe Ile His Glu Gly Tyr Thr His Asp Ala Gly Phe 225 230 235 240 Asp Asn Asp Ile Ala Leu Ile Lys Leu Asn Asn Lys Val Val Ile Asn 245 250 255 Ser Asn Ile Thr Pro Ile Cys Leu Pro Arg Lys Glu Ala Glu Ser Phe 260 265 270 Met Arg Thr Asp Asp Ile Gly Thr Ala Ser Gly Trp Gly Leu Thr Gln 275 280 285 Arg Gly Phe Leu Ala Arg Asn Leu Met Tyr Val Asp Ile Pro Ile Val 290 295 300 Asp His Gln Lys Cys Thr Ala Ala Tyr Glu Lys Pro Pro Tyr Pro Arg 305 310 315 320 Gly Ser Val Thr Ala Asn Met Leu Cys Ala Gly Leu Glu Ser Gly Gly 325 330 335 Lys Asp Ser Cys Arg Gly Asp Ala Gly Gly Ala Leu Val Phe Leu Asp 340 345 350 Ser Glu Thr Glu Arg Trp Phe Val Gly Gly Ile Val Ser Trp Gly Ser 355 360 365 Met Asn Cys Gly Glu Ala Gly Gln Tyr Gly Val Tyr Thr Lys Val Ile 370 375 380 Asn Tyr Ile Pro Trp Ile Glu Asn Ile Ile Ser Asp Phe 385 390 395 <210> 2 <211> 397 <212> PRT <213> Macaca fascicularis <400> 2 His His His His His His Ala Ser Gln Pro Cys Pro Tyr Pro Met Ala 1 5 10 15 Pro Pro Asn Gly His Leu Ser Pro Val Gln Ala Lys Tyr Ile Leu Lys 20 25 30 Asp Ser Phe Ser Ile Phe Cys Glu Pro Gly Tyr Glu Leu Leu Gln Gly 35 40 45 His Leu Pro Leu Lys Ser Phe Ala Ala Val Cys Gln Lys Asp Gly Ser 50 55 60 Trp Asp Gln Pro Met Pro Ser Cys Ser Ile Val Asp Cys Gly Pro Pro 65 70 75 80 Asp Asp Leu Pro Ser Gly Arg Val Glu Tyr Ile Thr Gly Pro Glu Val 85 90 95 Thr Thr Tyr Lys Ala Val Ile Gln Tyr Ser Cys Glu Glu Thr Phe Tyr 100 105 110 Thr Met Lys Val Asn Asp Gly Lys Tyr Val Cys Glu Ala Asp Gly Phe 115 120 125 Trp Thr Ser Ser Lys Gly Glu Arg Ser Pro Pro Val Cys Glu Pro Val 130 135 140 Cys Gly Leu Ser Ala Arg Thr Thr Gly Gly Arg Ile Tyr Gly Gly Gln 145 150 155 160 Lys Ala Lys Pro Gly Asp Phe Pro Trp Gln Val Leu Ile Leu Gly Gly 165 170 175 Ser Thr Ala Ala Gly Ala Leu Leu Tyr Asp Asn Trp Val Leu Thr Ala 180 185 190 Ala His Ala Ile Tyr Glu Gln Lys His Asp Ala Ser Ser Leu Asp Ile 195 200 205 Arg Leu Gly Ala Leu Lys Arg Leu Ser Pro His Tyr Thr Gln Ala Trp 210 215 220 Ala Glu Ala Val Phe Ile His Glu Gly Tyr Thr His Asp Ala Gly Phe 225 230 235 240 Asp Asn Asp Ile Ala Leu Ile Lys Leu Asn Asn Lys Val Val Ile Asn 245 250 255 Ser Asn Ile Thr Pro Ile Cys Leu Pro Arg Lys Glu Ala Glu Ser Phe 260 265 270 Met Arg Thr Asp Asp Ile Gly Thr Ala Ser Gly Trp Gly Leu Thr Gln 275 280 285 Arg Gly Leu Leu Ala Arg Asn Leu Met Tyr Val Asp Ile Pro Ile Val 290 295 300 Asp His Gln Lys Cys Thr Ala Ala Tyr Glu Lys Pro Pro Tyr Ser Gly 305 310 315 320 Gly Ser Val Thr Ala Asn Met Leu Cys Ala Gly Leu Glu Ser Gly Gly 325 330 335 Lys Asp Ser Cys Arg Gly Asp Ala Gly Gly Ala Leu Val Phe Leu Asp 340 345 350 Asn Glu Thr Gln Arg Trp Phe Val Gly Gly Ile Val Ser Trp Gly Ser 355 360 365 Met Asn Cys Gly Glu Ala Gly Gln Tyr Gly Val Tyr Thr Lys Val Ile 370 375 380 Asn Tyr Ile Pro Trp Ile Lys Asn Ile Ile Ser Asn Phe 385 390 395 <210> 3 <211> 396 <212> PRT <213> Mus musculus <400> 3 His His His His His His Ala Ser Arg Pro Cys Pro Asp Pro Thr Ala 1 5 10 15 Pro Pro Asn Gly Ser Ile Ser Pro Val Gln Ala Ile Tyr Val Leu Lys 20 25 30 Asp Arg Phe Ser Val Phe Cys Lys Thr Gly Phe Glu Leu Leu Gln Gly 35 40 45 Ser Val Pro Leu Lys Ser Phe Thr Ala Val Cys Gln Lys Asp Gly Ser 50 55 60 Trp Asp Arg Pro Met Pro Glu Cys Ser Ile Ile Asp Cys Gly Pro Pro 65 70 75 80 Asp Asp Leu Pro Asn Gly His Val Asp Tyr Ile Thr Gly Pro Glu Val 85 90 95 Thr Thr Tyr Lys Ala Val Ile Gln Tyr Ser Cys Glu Glu Thr Phe Tyr 100 105 110 Thr Met Ser Ser Asn Gly Lys Tyr Val Cys Glu Ala Asp Gly Phe Trp 115 120 125 Thr Ser Ser Lys Gly Glu Lys Leu Pro Pro Val Cys Glu Pro Val Cys 130 135 140 Gly Leu Ser Thr His Thr Ile Gly Gly Arg Ile Val Gly Gly Gln Pro 145 150 155 160 Ala Lys Pro Gly Asp Phe Pro Trp Gln Val Leu Leu Leu Gly Gln Thr 165 170 175 Thr Ala Ala Ala Gly Ala Leu Ile His Asp Asn Trp Val Leu Thr Ala 180 185 190 Ala His Ala Val Tyr Glu Lys Arg Met Ala Ala Ser Ser Leu Asn Ile 195 200 205 Arg Met Gly Ile Leu Lys Arg Leu Ser Pro His Tyr Thr Gln Ala Trp 210 215 220 Pro Glu Glu Ile Phe Ile His Glu Gly Tyr Thr His Gly Ala Gly Phe 225 230 235 240 Asp Asn Asp Ile Ala Leu Ile Lys Leu Lys Asn Lys Val Thr Ile Asn 245 250 255 Gly Ser Ile Met Pro Val Cys Leu Pro Arg Lys Glu Ala Ala Ser Leu 260 265 270 Met Arg Thr Asp Phe Thr Gly Thr Val Ala Gly Trp Gly Leu Thr Gln 275 280 285 Lys Gly Leu Leu Ala Arg Asn Leu Met Phe Val Asp Ile Pro Ile Ala 290 295 300 Asp His Gln Lys Cys Thr Ala Val Tyr Glu Lys Leu Tyr Pro Gly Val 305 310 315 320 Arg Val Ser Ala Asn Met Leu Cys Ala Gly Leu Glu Thr Gly Gly Lys 325 330 335 Asp Ser Cys Arg Gly Asp Ala Gly Gly Ala Leu Val Phe Leu Asp Asn 340 345 350 Glu Thr Gln Arg Trp Phe Val Gly Gly Ile Val Ser Trp Gly Ser Ile 355 360 365 Asn Cys Gly Ala Ala Asp Gln Tyr Gly Val Tyr Thr Lys Val Ile Asn 370 375 380 Tyr Ile Pro Trp Ile Glu Asn Ile Ile Ser Asn Phe 385 390 395 <210> 4 <211> 396 <212> PRT <213> Rattus norvegicus <400> 4 His His His His His His Ala Ser Gln Pro Cys Pro Asp Pro Thr Ala 1 5 10 15 Pro Pro Asn Gly His Ile Ser Pro Val Gln Ala Thr Tyr Val Leu Lys 20 25 30 Asp Ser Phe Ser Val Phe Cys Lys Thr Gly Phe Glu Leu Leu Gln Gly 35 40 45 Ser Val Pro Leu Lys Ser Phe Thr Ala Val Cys Gln Lys Asp Gly Ser 50 55 60 Trp Asp Arg Pro Ile Pro Glu Cys Ser Ile Ile Asp Cys Gly Pro Pro 65 70 75 80 Asp Asp Leu Pro Asn Gly His Val Asp Tyr Ile Thr Gly Pro Glu Val 85 90 95 Thr Thr Tyr Lys Ala Val Ile Gln Tyr Ser Cys Glu Glu Thr Phe Tyr 100 105 110 Thr Met Ser Ser Asn Gly Lys Tyr Val Cys Glu Ala Asp Gly Phe Trp 115 120 125 Thr Ser Ser Lys Gly Glu Lys Ser Leu Pro Val Cys Lys Pro Val Cys 130 135 140 Gly Leu Ser Thr His Thr Ser Gly Gly Arg Ile Ile Gly Gly Gln Pro 145 150 155 160 Ala Lys Pro Gly Asp Phe Pro Trp Gln Val Leu Leu Leu Gly Glu Thr 165 170 175 Thr Ala Ala Gly Ala Leu Ile His Asp Asp Trp Val Leu Thr Ala Ala 180 185 190 His Ala Val Tyr Gly Lys Thr Glu Ala Met Ser Ser Leu Asp Ile Arg 195 200 205 Met Gly Ile Leu Lys Arg Leu Ser Leu Ile Tyr Thr Gln Ala Trp Pro 210 215 220 Glu Ala Val Phe Ile His Glu Gly Tyr Thr His Gly Ala Gly Phe Asp 225 230 235 240 Asn Asp Ile Ala Leu Ile Lys Leu Lys Asn Lys Val Thr Ile Asn Arg 245 250 255 Asn Ile Met Pro Ile Cys Leu Pro Arg Lys Glu Ala Ala Ser Leu Met 260 265 270 Lys Thr Asp Phe Val Gly Thr Val Ala Gly Trp Gly Leu Thr Gln Lys 275 280 285 Gly Phe Leu Ala Arg Asn Leu Met Phe Val Asp Ile Pro Ile Val Asp 290 295 300 His Gln Lys Cys Ala Thr Ala Tyr Thr Lys Gln Pro Tyr Pro Gly Ala 305 310 315 320 Lys Val Thr Val Asn Met Leu Cys Ala Gly Leu Asp Arg Gly Gly Lys 325 330 335 Asp Ser Cys Arg Gly Asp Ala Gly Gly Ala Leu Val Phe Leu Asp Asn 340 345 350 Glu Thr Gln Arg Trp Phe Val Gly Gly Ile Val Ser Trp Gly Ser Ile 355 360 365 Asn Cys Gly Gly Ser Glu Gln Tyr Gly Val Tyr Thr Lys Val Thr Asn 370 375 380 Tyr Ile Pro Trp Ile Glu Asn Ile Ile Asn Asn Phe 385 390 395 <210> 5 <211> 114 <212> PRT <213> Mus musculus <400> 5 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Ser Gly Asp Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu His Leu Ser Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Only Arg Tyr Trp Gly Asp Tyr Trp Gly His Gly Thr Thr Leu Thr Val 100 105 110 Ser Ser <210> 6 <211> 342 <212> DNA <213> Muscles <400> 6 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattg cctggactg gatccggcag 120 tttcaggag acaactgga gtggatgggc tacataagct acagtggttc cactagctat 180 aacccatctc tcaaagtcg aatctctatc accgagaca catccaagaa ccagttcttc 240 ctgcacttga gttctgtgac tactgaggac acagccacat atttctgtgc agatattgg 300 ggcgactact ggggccacgg caccactctc acagctct ca 342 <210> 7 <211> 7 <212> PRT <213> Muscles <400> 7 Thr Ser Asp Tyr Ala Trp Asn 1 5 <210> 8 <211> 16 <212> PRT <213> Muscles <400> 8 Tyr Ile Ser Tyr Ser Gly Ser Thr Ser Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 9 <211> 5 <212> PRT <213> Mus musculus <400> 9 Tyr Trp Gly Asp Tyr 1 5 <210> 10 <211> 107 <212> PRT <213> Mus musculus <400> 10 Asp Ser Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Val Thr Cys Lys Ala Ser His Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Phe Gln Gln Lys Pro Gly Gln Ser Pro Lys Gly Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Ser Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Leu Gln Tyr Asn Ser Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Val Lys 100 105 <210> 11 <211> 321 <212> DNA <213> Mus musculus <400> 11 gacagtgtga tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aggccagtca caatgtgggt actaatgtag cctggtttca acagaaacca 120 ggacaatctc ctaaaggact gatttactcg gcatcctacc ggtccagtgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagac ttcactctca ccatcagcaa tgtgcagtct 240 gaagacttgg cagagtattt ctgtctacaa tataacagca atcctctgac gttcggtgga 300 ggcaccaagc tggaggtcaa g 321 <210> 12 <211> 11 <212> PRT <213> Mus musculus <400> 12 Lys Ala Ser His Asn Val Gly Thr Asn Val Ala 1 5 10 <210> 13 <211> 7 <212> PRT <213> Mus musculus <400> 13 Dear Ala Dear Tyr Arg Dear Dear 1 5 <210> 14 <211> 9 <212> PRT <213> Mus musculus <400> 14 Leu Gln Tyr Asn Ser Asn Pro Leu Thr 1 5 <210> 15 <211> 118 <212> PRT <213> Mus musculus <400> 15 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Leu Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Ser Ser Ala Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Ile Ile Ser Thr Ala Val Asp Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 16 <211> 354 <212> DNA <213> Mus musculus <400> 16 60. gatgtacaac ttcaggagtc gggacctggc ctggtgagac cttctcagtc tctgtccctc acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag tttccagga acaaactgga gtggatgggc tacataagtt acagtggtcg cactagttac aacccctctc tcaaaagtcg actctctatc actcgagaca catccaaga ccagttcttc ctgcagttga attctgtgag ttctgcggac acagccacgt attactgtgc aattatttct acggcagtag actttgacta ctggggccaa ggcaccactc tcacagtctc ctca <210> 17 <211> 16 <212> PRT <213> Mus musculus <400> 17 Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 18 <211> 9 <212> PRT <213> Mus musculus <400> 18 Ile Ser Thr Ala Val Asp Phe Asp Tyr 1 5 <210> 19 <211> 108 <212> PRT <213> Mus musculus <400> 19 Gln Thr Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Leu Gly 1 5 10 15 Glu Arg Val Thr Met Thr Cys Thr Ala Ser Ser Ser Val Ser Ser Ser 20 25 30 Tyr Leu Leu Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Leu Trp 35 40 45 Ile Tyr Ser Thr Ser Lys Leu Ala Ser Gly Val Pro Val Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu 65 70 75 80 Ala Asp Asp Ala Ala Thr Tyr Tyr Cys Leu Gln Tyr Asn Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 20 <211> 324 <212> DNA <213> Mus musculus <400> 20 60. caaactgttc tcacccagtc tccagcaatc atgtctgcat ctctagggga acgggtcacc atgacctgca ctgccagctc aagtgtaagt tccagttact tgctctggta ccagcagaag ccaggatcct cccccaaact ctggatttat agcacatcca agttggcttc tggagtccca 180 gttcgcttca gtggcagtgg gtctgggacc tcttactctc tcacaatcag cagcatggag 240 gctgacgatg ctgccactta ttactgcctc cagtataatc gttccccgct cacgttcggt 300 gctgggacca agctggagct 324 <210> 21 <211> 12 <212> PRT <213> Mus musculus <400> 21 Thr Ala Ser Ser Ser Val Ser Ser Tyr Leu Leu Free Mp3 Download 1 5 10 <210> 22 <211> 7 <212> PRT <213> Mus musculus <400> 22 Ser Thr Ser Lys Leu Ala Ser 1 5 <210> 23 <211> 9 <212> PRT <213> Mus musculus <400> 23 Leu Gln Tyr Asn Arg Ser Pro Leu Thr 1 5 <210> 24 <211> 117 <212> PRT <213> Mus musculus <400> 24 Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Ala Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Phe Ile Ser Tyr Ser Gly Arg Thr Asn Tyr Asn Pro Pro Leu 50 55 60 Thr Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Arg Ser Tyr Asp Tyr Trp Phe Gly Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ala 115 <210> 25 <211> 351 <212> DNA <213> Mus musculus <400> 25 gaggtccagc tgcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 gcctgcactg tcactggcta ctcaatcacc agtgattatg cctggcactg gatccggcag 120 tttccaggaa acaaactgga gtggatgggc ttcataagct acagtggtag aactaactac 180 aacccacctc tcacaagtg aatctctatc actcgagaca catccaagaa ccagttcttc 240 ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagatcatat 300 gattactggt ttggttattg gggccaaggg actctggtca ctgtctctgc a 351 <210> 26 <211> 7 <212> PRT <213> Mus musculus <400> 26 Thr Ser Asp Tyr Ala Trp His 1 5 <210> 27 <211> 16 <212> PRT <213> Mus musculus <400> 27 Phe Ile Ser Tyr Ser Gly Arg Thr Asn Tyr Asn Pro Pro Leu Thr Ser 1 5 10 15 <210> 28 <211> 8 <212> PRT <213> Mus musculus <400> 28 Ser Tyr Asp Tyr Trp Phe Gly Tyr 1 5 <210> 29 <211> 112 <212> PRT <213> Mus musculus <400> 29 Asn Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Met Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Thr Val Ser Arg Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Val Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110 <210> 30 <211> 336 <212> DNA <213> Mus musculus <400> 30 aatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60 atctcttgca gatctagtca gagtcttgta catagtaatg gaaacaccta tttagaatgg 120 tacatgcaga aaccaggcca gtctccaaag ctcctgatct acacagtttc cagacgattt 180 tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240 agcagagtgg aggctgagga tctgggagtt tattattgct ttcaaggttc acatgttccg 300 gtcacgtcg gtgctgggac caagctggag ctgaaa 336 <210> 31 <211> 16 <212> PRT <213> Mus musculus <400> 31 Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 32 <211> 7 <212> PRT <213> Mus musculus <400> 32 Thr Val Ser Arg Arg Phe Ser 1 5 <210> 33 <211> 9 <212> PRT <213> Mus musculus <400> 33 Phe Gln Gly Ser His Val Pro Val Thr 1 5 <210> 34 <211> 117 <212> PRT <213> Mus musculus <400> 34 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Lys Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Gly Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu His Leu Gln Ser Val Thr Ile Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Ile Leu Gly Tyr Phe Ala Met Asp Tyr Trp Gly Gln Gly Thr Pro 100 105 110 Val Thr Val Ser Ser 115 <210> 35 <211> 351 <212> DNA <213> Mus musculus <400> 35 60. gatgtacaac ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaagtg gatccggcag tttccagga acaaactgga gtggatgggc tacataagtt acagtggcag cactagctac 240. aacccttctc tcaaaggtcg aatctctatc actcgagaca catccaaga ccagtttttc ctgcacttgc aatctgtgc tattgaggac acagccacct attactgtgc aatactaggt tactttgcta tggactactg ggggcaagga accccagtca ccgtctcctc a 351 <210> 36 <211> 7 <212> PRT <213> Mus musculus <400> 36 Thr Ser Asp Tyr Ala Trp Light 1 5 <210> 37 <211> 16 <212> PRT <213> Mus musculus <400> 37 Tyr Ile Ser Tyr Ser Gly Ser Thr Ser Tyr Asn Pro Ser Leu Lys Gly 1 5 10 15 <210> 38 <211> 8 <212> PRT <213> Mus musculus <400> 38 Leu Gly Tyr Phe Ala Met Asp Tyr 1 5 <210> 39 <211> 107 <212> PRT <213> Mus musculus <400> 39 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Val Asp 20 25 30 Val Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Lys Ser Leu Ile 35 40 45 Tyr Leu Ala Ser Asn Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Leu Gln His Trp Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 40 <211> 321 <212> DNA <213> Mus musculus <400> 40 gacattgtga tgacccagtc tcaaaaattc atgtccactt cagtaggaga cagggtcagc 60 atcacctgca aggccagtca gaatgttcgt gttgacgttg cctggtatca acagaaatca 120 gggcagtctc ctaaatcact gatttacttg gcatccaacc ggcacactgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactctca ccattagcaa tgtgcaatct 240 gaagacctgg cagattattt ctgtctacaa cattggagtt atcctctgac gttcggtgga 300 ggcaccaagc tggaaatcaa a 321 <210> 41 <211> 11 <212> PRT <213> Mus musculus <400> 41 Lys Ala Ser Gln Asn Val Arg Val Asp Val Ala 1 5 10 <210> 42 <211> 7 <212> PRT <213> Mus musculus <400> 42 Leu Ala Ser Asn Arg His Thr 1 5 <210> 43 <211> 9 <212> PRT <213> Mus musculus <400> 43 Leu Gln His Trp Ser Tyr Pro Leu Thr 1 5 <210> 44 <211> 118 <212> PRT <213> Mus musculus <400> 44 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Thr Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Ile Gly Arg Ile Ser Ile Thr Arg Asp Thr Thr Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Ser Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Lys Leu Ala Gly Gly His Gly Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Ser Val Thr Val Ser Ser 115 <210> 45 <211> 354 <212> DNA <213> Mus musculus <400> 45 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120 tttccaggaa acacattgga gtggatgggc tatataagtt atagtggtag gactagctac 180 aacccatctc tgataggtcg aatctcaatc actcgagaca caaccaagaa ccagttcttt 240 ctgcagttga attctgtgtc tactgaggac acagccacat attactgtgc aaaattggcc 300 ggaggacatg ggatggacta ctggggtcaa ggaacctcag tcaccgtctc ctca 354 <210> 46 <211> 16 <212> PRT <213> Mus musculus <400> 46 Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu Ile Gly 1 5 10 15 <210> 47 <211> 9 <212> PRT <213> Mus musculus <400> 47 Leu Ala Gly Gly His Gly Met Asp Tyr 1 5 <210> 48 <211> 106 <212> PRT <213> Mus musculus <400> 48 Asn Ile Val Met Thr Gln Ser Gln Gln Phe Met Ser Val Gly 1 5 10 15 Asp Arg Will Thr Cys Lys Only Be Gln Asn Will Asp Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Leu Gln Tyr Asn Ser Tyr Leu Thr 85 90 95 Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 49 <211> 318 <212> DNA <213> Muscles <400> 49 aacattgtga tgacccagtc tcaacaattc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aggccagtca gatgtggat actaatgtag cctggtatca acagaaacca 120 <h2 style=";text-align:left;direction:ltr">ggacaatctc ctaaagcact gatttactcg gcatcctacc ggtacagtgg agtccctgat 180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> cgcttcacag gcagtggatc tgggacagat ttcactctca ccatcagcaa tgtgcagtct 240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> gaagacttgg cagaatattt ctgtctgcaa tataatagtt acctcacgtt cggttctggg 300<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> accaagctgg agctgaaa 318<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 50<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 11<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 50<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Lys Ala Ser Gln Asn Val Asp Thr Asn Val Ala<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5 10<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 51<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 7<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 51<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ser Ala Ser Tyr Arg Tyr Ser<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 52<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 8<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 52<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Leu Gln Tyr Asn Ser Tyr Leu Thr<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 53<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 118<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 53<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Arg Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn With Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Leu Ser Val Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Asn Ala Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Only Val Asp Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser 115 <210> 54 <211> 354 <212> DNA <213> Muscles <400> 54 gatgtacaac ttcaggagtc gggacctggc ctggtgagac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattg cctggactg gatccggcag 120 <h2 style=";text-align:left;direction:ltr">tttccaggaa acaaactgga gtggatggggc tacataagtt acagtggtcg cactagctac 180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> aacccctctc tcaagagtcg actctctgtc actcgagaca catccaagaa ccagttcttc 240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ctgcagttga attctgtgaa tgctgcggac acagccacgt attactgtgc aattatttct 300<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> acggcagtag actttgacta ctggggccaa ggcaccactc tcacagtctc ctca 354<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 55<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 108<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 55<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala Ser Leu Gly<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5 10 15<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Glu Arg Val Thr Met Thr Cys Thr Ala Ser Ser Ser Val Ser Ser Ser<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 20 25 30<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Tyr Leu Leu Trp Tyr Gln Gln Lys Pro Gly Ser Ser Pro Lys Leu Trp<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 35 40 45<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ile Tyr Ser Ser Ser Lys Leu Ala Ser Gly Val Pro Val Arg Phe Ser<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 50 55 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 65 70 75 80<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Leu Gln Tyr Asn Arg Ser Pro 85 90 95 Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 56 <211> 324 <212> DNA <213> Mus musculus <400> 56 caaattgttc tcacccagtc tccagcaatc atgtctgcat ctctagggga acgggtcacc 60 atgacctgca ctgccagctc aagtgtaagt tccagttact tgctctggta ccagcagaag 120 ccaggatcct cccccaaact ctggatttat agctcatcca agttggcttc tggagtccca 180 gttcgcttca gtggcagtgg gtctgggacc tcttactctc tcacaatcag cagcatggag 240 gctgaagatg ctgccactta ttactgcctc cagtataatc gttccccgct cacgttcggt 300 gctgggacca agctggagct gaaa 324 <210> 57 <211> 7 <212> PRT <213> Mus musculus <400> 57 Dear Dear Dear Lys Leu Here Dear 1 5 <210> 58 <211> 118 <212> PRT <213> Mus musculus <400> 58 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Lys Leu Gly Gly Gly His Gly Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Ser Val Thr Val Ser Ser 115 <210> 59 <211> 354 <212> DNA <213> Mus musculus <400> 59 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 <h2 style=";text-align:left;direction:ltr">acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tttccaggaa acaaattgga gtggatggggc tacataagtt atagtggtag gactagctac 180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> aacccatctc tgaaaagtcg aatctcaatc actcgagaca catccaagaa ccagttcttc 240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aaaattggggc 300<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ggaggacatg ggatggacta ctggggtcaa ggaacctcag tcaccgtctc ctca 354<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 9<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Leu Gly Gly Gly His Gly Met Asp Tyr<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 61<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 105<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 61<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly Asp<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5 10 15<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Asp Thr Asn Val<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 20 25 30<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile Tyr<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 35 40 45<h2 style=";text-align:left;direction:ltr"> Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser Glu 65 70 75 80 Asp With Glu Tyr Phe Cys With Gln Tyr Asn Ser Tyr With Thr Phe 85 90 95 Gly Ser Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 62 <211> 317 <212> DNA <213> Muscles <400> 62 acatgtgat gacccagtct caaaaattca tgtccacatc agtaggagac agggtcagcg 60 tcacctgcaa ggccagtcag aatgtggata ctaatgtagc ctggtatcaa cagaaaccag 120 ggcaatctcc taaagcgctg atttactcgg catcctaccg ttacagtgga gtccctgatc 180 gcttcacagg cagtgatct gggacagat tcactcac catcagcaat gtgcagtctg 240 aagacttggc agagtatttc tgtctgcaat atacagcta cctcacgttc ggttctggga 300 ccaagctgga gctgaaa 317 <210> 63 <211> 120 <212> PRT <213> Mus musculus <400> 63 Gln Val Thr Leu Lys Glu Ser Gly Pro Gly Ile Leu Lys Pro Ser Gln 1 5 10 15 Thr Leu Ile Leu Thr Cys Ser Phe Ser Gly Phe Ser Leu Asn Thr Ser 20 25 30 Gly Met Gly Val Gly Trp Ile Arg Gln Pro Ser Gly Lys Gly Leu Glu 35 40 45 Trp Leu Ala His Ile Trp Trp Asp Asp Asp Lys Asp Tyr Asn Pro Ser 50 55 60 Leu Lys Ser Gln Val Thr Ile Ser Lys Asp Thr Ser Thr Asn Gln Val 65 70 75 80 Phe Leu Lys Ile Thr Ser Val Asp Thr Ala Asp Gly Ala Thr Tyr Tyr 85 90 95 Cys Val Arg Asp Thr Thr Phe Leu Asp Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 64 <211> 360 <212> DNA <213> Mus musculus <400> 64 caagttactc taaaagagtc tggccctggg atattgaagc cctcacagac cctcattctg 60 acttgttctt tctctgggtt ttcactgaac acttctggta tgggtgtagg ctggattcgt 120 cagccttcag ggaagggtct ggagtggctg gcacacattt ggtgggatga tgataaggac 180 tataacccat ccctgaagag ccaagtcaca atctccaagg atacctccac aaaccaggtg 240 ttcctcaaga tcaccagtgt ggacactgca gatggtgcca cttactactg tgttcgagat 300 actacgtttt tagatgctat ggactactgg ggtcaaggaa cctcagtcac cgtctcctca 360 <210> 65 <211> 8 <212> PRT <213> Mus musculus <400> 65 Asn Thr Ser Gly Met Gly Val Gly 1 5 <210> 66 <211> 16 <212> PRT <213> Mus musculus <400> 66 His Ile Trp Trp Asp Asp Asp Lys Asp Tyr Asn Pro Ser Leu Lys Ser 1 5 10 15 <210> 67 <211> 10 <212> PRT <213> Mus musculus <400> 67 Asp Thr Thr Phe Leu Asp Ala Met Asp Tyr 1 5 10 <210> 68 <211> 112 <212> PRT <213> Mus musculus <400> 68 Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Arg 20 25 30 Asn Gly Asn Thr Phe Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Phe Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 110 <210> 69 <211> 336 <212> DNA <213> Mus musculus <400> 69 gatgttttga tgacccaaac tccactctcc ctgcctgtca gtcttggaga tcaagcctcc 60 atctcttgca ggtctagtca gagcattgta catcgtaatg gaaacacctt tttagaatgg 120 tacctgcaga aaccaggcca gtctccaaag ctcctgatct acagggtttc caaccgattt 180 tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240 agcagagtgg aggctgagga tctgggagtt tattactgct ttcaaggttc acatgttcca 300 ttcacgtcg gctcggggac aaagttggaa ataaaa 336 <210> 70 <211> 16 <212> PRT <213> Mus musculus <400> 70 Arg Ser Ser Gln Ser Ile Val His Arg Asn Gly Asn Thr Phe Leu Glu 1 5 10 15 <210> 71 <211> 7 <212> PRT <213> Mus musculus <400> 71 Arg Val Ser Asn Arg Phe Ser 1 5 <210> 72 <211> 9 <212> PRT <213> Mus musculus <400> 72 Phe Gln Gly Ser His Val Pro Phe Thr 1 5 <210> 73 <211> 118 <212> PRT <213> Mus musculus <400> 73 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Leu Pro Gly Asn Lys Leu Glu Trp 35 40 45 Val Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Thr Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Lys Leu Ala Gly Gly His Gly Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Ser Val Thr Val Ser Ser 115 <210> 74 <211> 354 <212> DNA <213> Mus musculus <400> 74 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattg cctggactg gatccggcag 120 ttaccaggaa acaattgga gtgggtgggc tatataagtt atagtggtag gactacctac 180 aacccatctc tgaaaagtcg aatctcaatc actcgagaca caaccagaa ccagttctc 240 ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aaagttggcc 300 gggggacatg ggatggacta ctggggtca ggaacctcag tcacctctc ctca 354 <210> 75 <211> 106 <212> PRT <213> Muscles <400> 75 Asn Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Val Gly 1 5 10 15 Asp Arg Will Thr Cys Lys Only Be Gln Asn Will Asp Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 His Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Ile Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Leu Gln Tyr Asn Ser Tyr Leu Thr 85 90 95 Phe Gly Ser Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 76 <211> 318 <212> DNA <213> Mus musculus <400> 76 aacattgtga tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aggccagtca gaatgtggat actaatgtag cctggtatca acagaaacca 120 gggcaatctc ctaaagcact gattcactcg gcatcctacc ggtacagtgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcattctca ccatcagtaa tgtgcagtct 240 gaagacttgg cagagtattt ctgtctgcaa tataacagct acctcacgtt cggttctggg 300 accaagctgg agctgaaa 318 <210> 77 <211> 116 <212> PRT <213> Mus musculus <400> 77 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn With Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Ser Asn Gln Phe Phe 65 70 75 80 Leu His Leu Asn Ser Val Thr Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Val Leu Gly Leu Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Pro Leu 100 105 110 Thr Val Ser Ser 115 <210> 78 <211> 348 <212> DNA <213> Muscles <400> 78 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattg cctggactg gatccggcag 120 tttccaggaa acaactgga gtggatgggc tacataagct acagtggtag aactacctac 180 aacccatctc tcaaagtcg aatctctatc accgagaca catccagaa ccagttctc 240 ctgcacttga attctgtgac tactgaggac acagccacat atttctgtgc agtactggga 300 ctctactttg actactgggg cctctcacag tctcctca 348 <210> 79 <211> 7 <212> PRT <213> Muscles <400> 79 Leu Gly Leu Tyr Phe Asp Tyr 1 5 <210> 80 <211> 107 <212> PRT <213> Muscles <400> 80 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Will Thr Cys Lys Only Be Gln Asn Will Asp Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys His Gln Phe Asn Ser Asn Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 81 <211> 321 <212> DNA <213> Mus musculus <400> 81 gacattgtga tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aagccagtca gaatgtggat actaatgtag cctggtatca acagaaacca 120 gggcaatctc ctaaagcact gatttactcg gcatcctacc ggtacagtgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactctca ccatcagcaa tgtgcagtct 240 gaagacttgg cagaatatttt ctgtcaccaa tttaacagca atcctctcac gttcggtgct 300 gggaccaagc tggagctgaa a 321 <210> 82 <211> 9 <212> PRT <213> Mus musculus <400> 82 His Gln Phe Asn Ser Asn Pro Leu Thr 1 5 <210> 83 <211> 117 <212> PRT <213> Mus musculus <400> 83 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Lys Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Ser Tyr His Pro Ser Leu 50 55 60 Lys Gly Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Ala Asp Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Ile Leu Gly Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser 100 105 110 Val Thr Val Ser Ser 115 <210> 84 <211> 351 <212> DNA <213> Mus musculus <400> 84 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattg cctggaatg gatccggcag 120 tttccaggaa acaactgga atggatgggc tacataagct acagtggttc cactagctat 180 cacccatctc tcaaggtcg aatctccatc actcgagaca catccagaa ccagttctc 240 ctgcagttga attctgtgac tgctgacgac acagccacat attactgtgc atattaggt 300 tactatgcta tggactactg gggtcagga acctcagtca ccgtctccctc a 351 <210> 85 <211> 16 <212> PRT <213> Muscles <400> 85 Tyr Ile Ser Tyr Ser Gly Ser Thr Ser Tyr His Pro Ser Leu Lys Gly 1 5 10 15 <210> 86 <211> 8 <212> PRT <213> Muscles <400> 86 Leu Gly Tyr Tyr Ala Met Asp Tyr 1 5 <210> 87 <211> 107 <212> PRT <213> Muscles <400> 87 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Ile Thr Cys Lys Ala Ser Gln Asn Val Arg Val Asp 20 25 30 Val Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Arg Ala Leu Ile 35 40 45 Tyr Leu Ala Ser Asn Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Leu Gln His Trp Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 88 <211> 321 <212> DNA <213> Mus musculus <400> 88 gacattgtga tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagt 60 atcacctgca aggccagtca gaatgttcgt gttgacgtag cctggtatca acagaaatca 120 gggcagtctc ctagagcact gatttacttg gcatccaacc ggcacactgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactca ccattaccaa tgtgcaatct 240 gaggacctgg cagattattt ctgtctacaa cattggagtt atcctctgac gttcggtgga 300 321 ggcaccaagc tggaatcaa a 321 <210> 89 <211> 107 <212> PRT <213> Muscles <400> 89 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Will Have Thr Cys Lys Only Be Gln Asn Or Arg Will Asp 20 25 30 Val Ala Trp Tyr Gln Gln Lys Ser Gly Gln Ser Pro Arg Ala Leu Ile 35 40 45 Tyr Leu Ala Ser Asn Arg His Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Asp Tyr Phe Cys Pro Gln His Trp Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 90 <211> 321 <212> DNA <213> Mus musculus <400> 90 gacattgtga tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagt 60 atcacctgca aggccagtca gaatgttcgt gttgacgtag cctggtatca acagaaatca 120 gggcagtctc ctagagcact gatttacttg gcatccaacc ggcacactgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactctca ccattaccaa tgtgcaatct 240 gaggacctgg cagattattt ctgtccacaa cattggagtt atcctctgac gttcggtgga 300 ggcaccaagc tggaaatcaa a 321 <210> 91 <211> 9 <212> PRT <213> Mus musculus <400> 91 Pro Gln His Trp Ser Tyr Pro Leu Thr 1 5 <210> 92 <211> 119 <212> PRT <213> Mus musculus <400> 92 Lys Val Lys Leu Val Glu Ser Gly Gly Asp Leu Val Lys Pro Gly Gly 1 5 10 15 Leu Lys Ile Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Only Met Ser TRP With Arg Gln Thr Pro Asp Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Val Ser Ser Gly Gly Pro Tyr Thr Tyr Tyr Pro Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr With Arg Asp Asn Ala Glu Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Ser Leu Arg Ser Asp Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Only Arg Gln With Tyr Ser Asp Tyr Asp Val Thr Tyr Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr Val Ser Ala 115 <210> 93 <211> 357 <212> DNA <213> Muscles <400> 93 aaagtgaagc tggtggagtc tgggggac ttagtgaagc ctggaggtc cctgaaaatc 60 tcctgtacag cctctggatt cacttcagt aactagcca tgtcttggat tcgccagact 120 ccggacaaga ggctggagtg ggtcgcaacc gttagtagtg gtggtcctta cacctactat 180 ccagacagtg tgaaggggcg attcaccatc tccagagaca atgccgaaaa caccctgtac 240 ctgcaaatga gcagtctgag gtctgatgac acggccatat attactgtgc aaggcagatc 300 tactctgatt acgacgtcac ttactggggc caagggactc tggtcactgt ctctgca 357 <210> 94 <211> 6 <212> PRT <213> Mus musculus <400> 94 Ser Asn Tyr Ala Met Ser 1 5 <210> 95 <211> 17 <212> PRT <213> Mus musculus <400> 95 Thr Val Ser Ser Gly Gly Pro Tyr Thr Tyr Tyr Pro Asp Ser Val Lys 1 5 10 15 Gly <210> 96 <211> 10 <212> PRT <213> Mus musculus <400> 96 Gln Ile Tyr Ser Asp Tyr Asp Val Thr Tyr 1 5 10 <210> 97 <211> 106 <212> PRT <213> Mus musculus <400> 97 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Thr Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Phe Met 20 25 30 Tyr Trp Leu Gln Gln Lys Pro Gly Ser Ser Pro Gln Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Thr Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Phe Arg Pro Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 98 <211> 318 <212> DNA <213> Mus musculus <400> 98 caaattgttc tctcccagtc tccagcaatc ctgtctgcat ctccagggga gacggtcaca 60 atgacttgca gggccagttc aagtgtaagt ttcatgtact ggctccagca gaagccaggt 120 tcctcccccc aaccctggat tttgccaca tccaacctgg cttctggagt ccctgctcgc 180 ttcagtggca gtgggtctgg gacctcttac tccctcacaa tcaccagagt ggaggctgaa 240 gatgctgcca cttattactg tcaacagtgg agttttagac cgtggacgtt cggtggaggc 300 accaagctgg aaatcaaa 318 <210> 99 <211> 10 <212> PRT <213> Mus musculus <400> 99 Arg Ala Dear Dear Dear Val Dear Phe Met Tyr 1 5 10 <210> 100 <211> 7 <212> PRT <213> Mus musculus <400> 100 Ala Thr Sister Donkey Leu Ala Sister 1 5 <210> 101 <211> 9 <212> PRT <213> Mus musculus <400> 101 Gln Gln Trp Ser Phe Arg Pro Trp Thr 1 5 <210> 102 <211> 117 <212> PRT <213> Mus musculus <400> 102 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Phe Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Phe Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Arg Ser Tyr Asp Tyr Trp Phe Pro Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ala 115 <210> 103 <211> 351 <212> DNA <213> Mus musculus <400> 103 gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 <h2 style=";text-align:left;direction:ltr">acctgcactg tcactggctt ctcaatcacc agtgattatg cctggaactg gatccggcag 120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tttccaggaa acaaactgga gtggatggggc tacataagct acagtggtag aactaactac 180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> aacccatctc tcaaaagtcg attctctatc actcgagaca catccaagaa ccagttcttc 240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagatcatat 300<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> gattactggt ttccttactg gggccaaggg actctggtca ctgtctctgc a 351<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 104<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 16<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 104<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Tyr Ile Ser Tyr Ser Gly Arg Thr Asn Tyr Asn Pro Ser Leu Lys Ser<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5 10 15<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 105<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 8<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 105<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ser Tyr Asp Tyr Trp Phe Pro Tyr<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 106<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 112<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 106<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Asp Val Leu Met Thr Gln Thr Pro Leu Ser Leu Pro Val Ser Leu Gly<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 1 5 10 15 Asp Gln Ala Ser Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Phe Glu Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 His Ile Pro Val Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110 <210> 107 <211> 336 <212> DNA <213> Muscles <400> 107 gatgttttga tgacccaac tccactcc ctgcctgtca gtcttggaga tcaagcctcc 60 atctcttgca gatctagtca gagcattgta catagtaatg gaacacctt ttggaatgg 120 tacctgcaga aaccaggcca gtctccaag ctcctgatct acaaagtttc siaccgattt 180 tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcaagatc 240 agcagagtgg aggctgagga tctgggagtt tattactgct ttcaaggttc acatattccg 300 gtcacgttcg gtgctgggac caagctggag ctgaaa 336 <210> 108 <211> 16 <212> PRT <213> Mus musculus <400> 108 Arg Ser Ser Gln Ser Ile Val His Ser Asn Gly Asn Thr Phe Leu Glu 1 5 10 15 <210> 109 <211> 7 <212> PRT <213> Mus musculus <400> 109 Lys Val Ser Asn Arg Phe Ser 1 5 <210> 110 <211> 9 <212> PRT <213> Mus musculus <400> 110 Phe Gln Gly Ser His Ile Pro Val Thr 1 5 <210> 111 <211> 117 <212> PRT <213> Mus musculus <400> 111 Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Phe Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp His Trp With Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Phe Ile Asn Tyr Ser Gly Arg Thr Asn Tyr Asn Pro Ser Leu 50 55 60 Ser Ile Arg Ile Ser Ile Ile Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Serves Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Arg Ser Tyr Asp Phe Trp Phe Gly Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Would Thr Would Be a Wing 115 <210> 112 <211> 351 <212> DNA <213> Muscles <400> 112 gaggtccagc tgcaggagtc gggacctgc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggctt ctcaatcacc agtgattg cctgcactg gatccggcag 120 tttccaggaa acaactgga gtggatgggc ttcataact acagtgtag aactactac 180 aacccatctc tcacaagtcg aatctctatc attcgggaca catccaagaa ccagttctc 240 ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagatcatat 300 gatttctggt ttggttattg gggccaaggg actctggtca ctgtctctgc a 351 <210> 113 <211> 16 <212> PRT <213> Muscles <400> 113 Phe Ile Asn Tyr Ser Gly Arg Thr Asn Tyr Asn Pro Ser Leu Thr Ser 1 5 10 15 <210> 114 <211> 8 <212> PRT <213> Muscles <400> 114 Ser Tyr Asp Phe Trp Phe Gly Tyr 1 5 <210> 115 <211> 112 <212> PRT <213> Muscles <400> 115 Asn Val Leu Met Thr Gln Ser Pro Leu Ser Leu Pro Val Ser Leu Gly 1 5 10 15 Asp Gln Ala Ser Ser Ile Ser Cys Arg Ser Ser Gln Ser Ile Val His Ser 20 25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Met Gln Lys Pro Gly Gln Ser 35 40 45 Pro Lys Leu Leu Ile Tyr Thr Val Ser Lys Arg Phe Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70 75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85 90 95 Ser His Val Pro Val Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 110 <210> 116 <211> 336 <212> DNA <213> Muscles <400> 116 aatgttttga tgacccaag tccactctcc ctgctgtca gtcttggaga tcaagcctcc 60 atctcttgca gatctagtca gagtattgta catagtaatg gaacaccta tttagaatgg 120 tatatgcaga aaccaggcca gtctccaaag ctcctgatct accagttc caaacgattt 180 tctggggtcc cagacaggtt cagtggcagt ggatcaggga cagatttcac actcagatc 240 agcagagtgg aggctgagga tctgggagtt tattattgct ttcaaggttc acatgttccg 300 gtcacgtcg gtgctgggac caagctggag ctgaaa 336 <210> 117 <211> 16 <212> PRT <213> Mus musculus <400> 117 Arg Ser Ser Gln Ser Ile Val His Ser Asn Gly Asn Thr Tyr Leu Glu 1 5 10 15 <210> 118 <211> 7 <212> PRT <213> Mus musculus <400> 118 Thr Val Ser Lys Arg Phe Ser 1 5 <210> 119 <211> 119 <212> PRT <213> Mus musculus <400> 119 Lys Val Lys Leu Val Glu Ser Gly Gly Asp Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Lys Ile Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30 Ala Met Ser Trp Ile Arg Gln Thr Pro Asp Lys Arg Leu Glu Trp Val 35 40 45 Ala Thr Val Ser Ser Gly Gly Pro Tyr Thr Tyr Tyr Pro Asp Ser Val<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 50 55 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Glu Asn Thr Leu Tyr<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 65 70 75 80<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Leu Gln Met Ser Ser Leu Arg Ser Asp Asp Thr Ala Ile Tyr Tyr Cys<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 85 90 95<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ala Arg Gln Ile Tyr Ser Asp Tyr Asp Val Thr Tyr Trp Gly Gln Gly<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 100 105 110<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Thr Leu Val Thr Val Ser Ala<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 115<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 357<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> DNA<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> aaagtgaagc tggtggagtc tgggggagac ttagtgaagc ctggagggtc cctgaaaatc 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tcctgtgcag cctctggatt cactttcagt aactatgcca tgtcttggat tcgccagact 120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ccggacaaga ggctggagtg ggtcgcaacc gttagtagtg gtggtcctta cacctactat 180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ccagacagtg tgaaggggcg attcaccatc tccagagaca atgccgaaaa caccctgtac 240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ctgcaaatga gcagtctgag gtctgatgac acggccattt attactgtgc aaggcagatc 300<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tactctgatt acgacgtcac ttactggggc caagggactc tggtcactgt ctctgca 357<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 121 <211> 106 <212> PRT <213> Mus musculus <400> 121 Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly 1 5 10 15 Glu Thr Val Thr Met Thr Cys Arg Val Ser Ser Ser Val Ser Tyr Met 20 25 30 Tyr Trp Leu Gln Gln Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr 35 40 45 Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Thr Arg Val Glu Ala Glu 65 70 75 80 Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Phe Arg Pro Trp Thr 85 90 95 Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 122 <211> 318 <212> DNA <213> Mus musculus <400> 122 caaattgttc tctcccagtc tccagcaatc ctgtctgcat ctccagggga gacggtcaca 60 atgacttgca gggtcagttc aagtgtaagt tacatgtact ggctccaaca gaagccagga 120 tcctccccca aaccctggat ttgccaca tccaacctgg cttctggagt ccctgctcgc 180 ttcagtggca gtggtctgg gacctac tcctcacaa tcaccagt ggaggctgaa 240 gatgctgcca cttattactg tcagcagtgg agttttagac cgtggacgtt cggtggaggc 300 accaagctgg aaatcaa 318 <210> 123 <211> 10 <212> PRT <213> Muscles <400> 123 Arg Will Be Ser Will Be Tyr Met Tyr 1 5 10 <210> 124 <211> 120 <212> PRT <213> Muscles <400> 124 Gln Val Gln Leu Gln Gln Ser Gly Thr Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Arg Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg Ser His 20 25 30 Thr Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Containing Pro Ser Thr Gly Tyr Containing Tyr Asn Gln Lys Phe 50 55 60 Lys Asp Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Only Arg Ser Asn Tyr Ser Tyr Tyr Tyr Gly Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 125 <211> 360 <212> DNA <213> Muscles <400> 125 caggtccagc tgcagcagtc tgggactgaa ctggcagac ctggggcctc agtgaggatg 60 tcctgcagg cttctggcta cacctttagg agtcacacga tgcactgggt aaaacagagg 120 cctggacagg gtctggaatg gattgatac attaatccta gcactggtta tattattac 180 aatcagaagt tcaaggacaa ggccacattg actgcagaca aatcctccag cacagcctac 240 atgcaactga gcagcctgac atctgaggac tctgcagtct attackgtgc aagaagtaac 300 tactcttatt actatggtat ggactactgg ggtcaaggaa cctcagtcac cgtctcctca 360 <210> 126 <211> 5 <212> PRT <213> Mus musculus <400> 126 Ser His Thr Met His 1 5 <210> 127 <211> 17 <212> PRT <213> Mus musculus <400> 127 Tyr Ile Asn Pro Ser Thr Gly Tyr Ile Asn Tyr Asn Gln Lys Phe Lys 1 5 10 15 Asp <210> 128 <211> 11 <212> PRT <213> Mus musculus <400> 128 Ser Asn Tyr Ser Tyr Tyr Tyr Gly Met Asp Tyr 1 5 10 <210> 129 <211> 107 <212> PRT <213> Mus musculus <400> 129 Asp Ile Val Met Thr Gln Ser His Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Asn Thr Ala 20 25 30 Can Only Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu Gln Ala 65 70 75 80 Glu Asp Leu Ala Val Tyr Tyr Cys Gln Gln His Tyr Asn Ile Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 130 <211> 321 <212> DNA <213> Muscles <400> 130 gatattgtga tgacccagtc tcacaattc atgtccacat cagtgggaga cagggtcacc 60 atcacctgca aggccagtca ggatgtaaat actgctgtag cctggtatca acagaaacca 120 ggacaatctc ctaaactact gatttactcg gcatcctacc ggtacactgg ggtccctgat 180 cgcttcactg gcagtgatc tgggacgat ttcacttca ccatcagcag tttacaggct 240 gaagacctgg cagtttatta ctgtcagcaa cattatata ttccgctcac gttcggtgct 300 gggaccaagc tggagctgaa a 321 <210> 131 <211> 11 <212> PRT <213> Mus musculus <400> 131 Lys Ala Ser Gln Asp Val Asn Thr Ala Val Ala 1 5 10 <210> 132 <211> 7 <212> PRT <213> Mus musculus <400> 132 Ser Ala Ser Tyr Arg Tyr Thr 1 5 <210> 133 <211> 9 <212> PRT <213> Mus musculus <400> 133 Gln Gln His Tyr Asn Ile Pro Leu Thr 1 5 <210> 134 <211> 120 <212> PRT <213> Mus musculus <400> 134 Gln Val Gln Leu Gln Gln Ser Gly Thr Glu Leu Ala Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Thr Tyr 20 25 30 Ser Ile His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Arg Asp Lys With Thr Leu Thr With Asp Arg Ser With Asn Thr With Tyr 65 70 75 80 Met Gln Leu Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Tyr Tyr Thr Phe Tyr His Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 135 <211> 360 <212> DNA <213> Muscles <400> 135 caggtccagc tgcagcagtc tgggactgaa ctggcagac ctggggcctc attgaagagatg 60 tcctgcagg cttctggcta cacctttgct acctactcga tacactgggt aaaacagagg 120 cctgggcagg gtctggaatg gattgatac attaatccta gcagtggtta tactattac 180 aatcagaagt tcagggacaa ggccacattg actgcagaca gatcctccaa cacagcctac 240 atgcaactga gcagcctgac atctgaggac tctgcagtct attackgtgc aagagtctac 300 tatacttttt accatgctat ggactactgg ggtcaaggaa cctcagtcac cgtctcctca 360 <210> 136 <211> 6 <212> PRT <213> Mus musculus <400> 136 Ala Thr Tyr Ser Ile His 1 5 <210> 137 <211> 17 <212> PRT <213> Mus musculus <400> 137 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Arg 1 5 10 15 Asp <210> 138 <211> 11 <212> PRT <213> Mus musculus <400> 138 Val Tyr Tyr Thr Phe Tyr His Ala Met Asp Tyr 1 5 10 <210> 139 <211> 114 <212> PRT <213> Mus musculus <400> 139 Gln Met Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn With Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Ser Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Serves Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Only Arg His Tyr Gly Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100 105 110 Ser Ser <210> 140 <211> 342 <212> DNA <213> Muscles <400> 140 cagatgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgcactg tcactggcta ctcaatcacc agtgattg cctggactg gatccggcag 120 tttccaggaa acaactgga gtggatgggc tacataagct acagtggtag gactacctac 180 aacccatctc tcaaagtcg aatctctatc accgagaca catccagaa ccagttctc 240 ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagacactac 300 ggagactact ggggccaagg caccactctc acagtctcct ca 342 <210> 141 <211> 5 <212> PRT <213> Mus musculus <400> 141 His Tyr Gly Asp Tyr 1 5 <210> 142 <211> 107 <212> PRT <213> Mus musculus <400> 142 Asp Ile Val Met Thr Gln Ser Gln Lys Leu Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Asn Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu Ile Lys 100 105 <210> 143 <211> 321 <212> DNA <213> Mus musculus <400> 143 gacattgtga tgactcagtc tcaaaaactc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aggccagtca gaatgtgggt actaatgtag cctggtatca acagaaacca 120 gggcaatctc ctaaagcact gatttactcg gcatcctacc ggaacagtgg agtccctgat 180 cgcttcacag gcagtggttc tgggacagat ttcactctca ccatcagcaa tgtgcagtct 240 gaagacttg cagagtattt ctgtcagcaa tttaacacca atcctctcac gttcggctcg 300 gggacaaagt tggaaataaa a 321 <210> 144 <211> 11 <212> PRT <213> Mus musculus <400> 144 Lys Ala Ser Gln Asn Val Gly Thr Asn Val Ala 1 5 10 <210> 145 <211> 7 <212> PRT <213> Mus musculus <400> 145 Ser Ala Ser Tyr Arg Asn Ser 1 5 <210> 146 <211> 9 <212> PRT <213> Mus musculus <400> 146 Gln Gln Phe Asn Thr Asn Pro Leu Thr 1 5 <210> 147 <211> 123 <212> PRT <213> Mus musculus <400> 147 Gln Val Gln Leu Gln Gln Ser Gly Pro Gln Leu Leu Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Phe Ser Phe Thr Asn His 20 25 30 Trp Ile His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Met Ile Asp Pro Ser Asp Ser Glu Thr Gly Leu Asn Gln Lys Phe 50 55 60 Arg Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 65 70 75 80 Met His Leu Asn Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Gly Ile His Asp Gly Tyr Tyr Ile Tyr Ala Met Asp Phe 100 105 110 Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser 115 120 <210> 148 <211> 369 <212> DNA <213> Mus musculus <400> 148 caggttcagc tgcagcagtc tgggcctcag ctacttaggc ctggggcttc agtgaagata 60 tcctgcaagg cttctggttt ccattcacc aaccactgga tacactgggt gaaacagagg 120 cctggacaag gtcttgagtg gattggcatg attgatcctt ccgatagtga aactggcctc 180 aatcagaagt tcagggacaa ggccacattg actgtagaca agtcctccag cacagcctac 240 atgcacctca acagcccgac atctgaggac tctgcggtct attactgtgc aagattggga 300 atccatgatg gttactacat ctatgctatg gacttttggg gtcaaggaac ctcagtcacc 360 gtctcctca 369 <210> 149 <211> 6 <212> PRT <213> Mus musculus <400> 149 Thr Asn His Trp Ile His 1 5 <210> 150 <211> 17 <212> PRT <213> Mus musculus <400> 150 Met Ile Asp Pro Ser Asp Ser Glu Thr Gly Leu Asn Gln Lys Phe Arg 1 5 10 15 Asp <210> 151 <211> 14 <212> PRT <213> Mus musculus <400> 151 Leu Gly Ile His Asp Gly Tyr Tyr Ile Tyr Ala Met Asp Phe 1 5 10 <210> 152 <211> 107 <212> PRT <213> Mus musculus <400> 152 Asn Ile Val Met Thr Gln Ser Pro Lys Ser Met Ser Met Ser Val Gly 1 5 10 15 Glu Arg Val Thr Leu Ser Cys Lys Ala Ser Glu Asn Val Gly Thr Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Lys Pro Glu Gln Ser Pro Lys Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Asn Arg Tyr Thr Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Ala Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala 65 70 75 80 Glu Asp Leu Ala Asp Tyr His Cys Gly Gln Ser Tyr Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 153 <211> 321 <212> DNA <213> Mus musculus <400> 153 aacattgtaa tgacccaatc tcccaaatcc atgtccatgt cagtaggaga gagggtcacc 60 ttgagctgca aggccagtga gaatgtgggt acttatgtat cctggtatca acagaaacca 120 gagcagtctc ctaaactgct gatatacggg gcatccaacc ggtacactgg ggtccccgat 180 cgcttcacag gcagtggatc tgcaacagat ttcactctga ccatcagcag tgtgcaggct 240 gaagaccttg cagattatca ctgtggacag agttacagct atccgctcac gttcggtgct 300 gggaccaagc tggagctgaa a 321 <210> 154 <211> 11 <212> PRT <213> Mus musculus <400> 154 Lys Ala Ser Glu Asn Val Gly Thr Tyr Val Ser 1 5 10 <210> 155 <211> 7 <212> PRT <213> Mus musculus <400> 155 Gly Ala Ser Asn Arg Tyr Thr 1 5 <210> 156 <211> 9 <212> PRT <213> Mus musculus <400> 156 Gly Gln Ser Tyr Ser Tyr Pro Leu Thr 1 5 <210> 157 <211> 114 <212> PRT <213> Mus musculus <400> 157 Glu Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Ser Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Ile Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Only Arg Tyr Trp Gly Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100 105 110 Ser Ser <210> 158 <211> 342 <212> DNA <213> Muscles <400> 158 gaggtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60 acctgctctg tcactggcta ctcaatcacc agtgattg cctggactg gatccggcag 120 tttccaggaa acagctgga gtggatgggc tacataagtt acagtggtag acactactac 180 aacccatctc tcaaagtcg aatctctatc accgagaca catccaagaa ccagttcttc 240 ctgcagttga tttctgtgac tactgaggac acagccacat attattgtgc agatattgg 300 ggcgactact ggggccaagg caccactctc acagctct ca 342 <210> 159 <211> 107 <212> PRT <213> Muscles <400> 159 Asp Ile Met Thr Gln Ser Gln Lys Phe Ile Thr Ser Ile Gly 1 5 10 15 Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asp Val Gly Gly His 20 25 30 Val Ala Trp Phe Gln Gln Arg Pro Gly Gln Ser Pro Lys Pro Leu Ile 35 40 45 Tyr Ser Ala Thr Phe Arg His Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Leu Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Val Cys Leu Gln Tyr Asn Ser Asn Pro Leu 85 90 95 Thr Phe Gly Pro Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 160 <211> 321 <212> DNA <213> Mus musculus <400> 160 gacattgtga tgacccagtc tcaaaaattc atctccacat caataggaga cagggtcagc 60 gtcacctgca aggccagtca ggatgtgggt ggacatgtgg cctggtttca acagagaccc 120 gggcaatctc ctaaacctct gatttattcg gcaaccttcc ggcacagtgg agtccctgat 180 cgcttcacgg gcagtggatc tgggacagat ttcactctca ccatcagtaa tctacagtct 240 gaggacttgg cagaatatgt ctgtctgcaa tataacagta atccactcac gttcggtcct 300 gggaccaagc tggagctgaa a 321 <210> 161 <211> 11 <212> PRT <213> Mus musculus <400> 161 Lys Ala Ser Gln Asp Val Gly Gly His Val Ala 1 5 10 <210> 162 <211> 7 <212> PRT <213> Mus musculus <400> 162 Ser Ala Thr Phe Arg His Ser 1 5 <210> 163 <211> 114 <212> PRT <213> Mus musculus <400> 163 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Tyr Leu 50 55 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 65 70 75 80<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 85 90 95<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ala Arg Tyr Trp Gly Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> 100 105 110<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> Ser Ser<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 164<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 342<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> DNA<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <213> Mus musculus<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <400> 164<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc 60<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag 120<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> tttccaggaa acaaactgga gtggatggggc tacataagct acagtggtcg cactagctac 180<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> aacccatatc tcaaaagtcg aatttctatc actcgagaca catccaagaa ccagttcttc 240<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagatattgg 300<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> ggcgactact ggggccaagg caccactctc acagtctcct ca 342<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <210> 165<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <211> 16<h2 style=";text-align:left;direction:ltr"> <h2 style=";text-align:left;direction:ltr"> <212> PRT<h2 style=";text-align:left;direction:ltr"> <213> Mus musculus <400> 165 Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Tyr Leu Lys Ser 1 5 10 15 <210> 166 <211> 107 <212> PRT <213> Mus musculus <400> 166 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Lys Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg His Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Leu Gln Tyr Asn Ser Asn Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 167 <211> 321 <212> DNA <213> Mus musculus <400> 167 gacattgtga tgacccagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aggccagtca gaatgtgggt actaatgtag cctggtataa acagaaacca 120 gggcaatctc ctaaagcact gatttactcg gcatcctacc ggcacagtgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactctca ccatcagcaa tgtgcagtct 240 gaagacttgg cagagtattt ctgtctgcaa tataacagca atcctctcac gttcggtgct 300 gggaccaagc tggagctgaa a 321 <210> 168 <211> 7 <212> PRT <213> Mus musculus <400> 168 Dear Here Dear Tyr Arg His Dear 1 5 <210> 169 <211> 114 <212> PRT <213> Mus musculus <400> 169 Asp Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1 5 10 15 Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40 45 Met Gly Thyr Ile Ser Thyr Ser Gly Arg Thr Ser Thyr Asn Pro Ser Leu 50 55 60 Light Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Light Asn Gln Phe Phe 65 70 75 80 Leu Gln Leu Asn Ser Val Thr Ala Glu Asp Thr Ala Thr Tyr Tyr Cys 85 90 95 Ala Arg Tyr Trp Gly Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100 105 110 Ser Ser <210> 170 <211> 342 <212> DNA <213> Mus musculus <400> 170 60. gatgtgcagc ttcaggagtc gggacctggc ctggtgaaac cttctcagtc tctgtccctc acctgcactg tcactggcta ctcaatcacc agtgattatg cctggaactg gatccggcag tttccagga acaaactgga gtggatgggc tacataagct acagtggtcg cactagctac 240. aacccatctc tcaaaagtcg aatctctatc actcgagaca catccaaga ccagttcttc ctgcagttga attctgtgac tgctgaggac acagccacat attactgtgc agatattgg 300 ggcgactact ggggccaagg caccactctc acagctct ca 342 <210> 171 <211> 107 <212> PRT <213> Muscles <400> 171 Asp Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly 1 5 10 15 Asp Arg Will Thr Cys Lys Only Be Gln Asn 20 25 30 Val Ala Trp Phe Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Thr Ser Tyr Arg Ser Gly Val Pro Asp Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn Val Gln Ser 65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Phe Asn Ser Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Leu Lys 100 105 <210> 172 <211> 321 <212> DNA <213> Mus musculus <400> 172 gacattgtga tgacacagtc tcaaaaattc atgtccacat cagtaggaga cagggtcagc 60 gtcacctgca aggccagtca gaatgtgggt actaatgtag cctggtttca acagaaacca 120 gggcaatctc ctaaagcact gatttactcg acatcctacc ggtccagtgg agtccctgat 180 cgcttcacag gcagtggatc tgggacagat ttcactctca ccatcagcaa tgtgcagtct 240 gaagacttgg cagagtactt ctgtcagcaa tttaacagca atcctctcac gttcggaggg 300 gggaccaagc tggagctgaa a 321 <210> 173 <211> 7 <212> PRT <213> Mus musculus <400> 173 Create Thr Create Tyr Arg Create Create 1 5 <210> 174 <211> 9 <212> PRT <213> Mus musculus <400> 174 Gln Gln Phe Asn Ser Asn Pro Leu Thr 1 5 <210> 175 <211> 114 <212> PRT <213> Homo sapiens <400> 175 Gln Met Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg His Tyr Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 100 105 110 Ser Ser <210> 176 <211> 342 <212> DNA <213> Homo sapiens <400> 176 cagatgcagc tgcaagagtc tggacctgga ctggtcaagc ccagcgagac actgagcctg 60 acctgtaccg tgtccggcta cagcatcaca agcgactacg cctggaactg gatcagacag 120 cctcctggca aaggcctcga gtggatcggc tacatcagct acagcggcag aaccacctac 180 aaccccagcc tgaagtccag agtgaccatc agcagagaca ccagcaagaa ccagttctcc 240 ctgaagctga gcagcgtgac agccgccgat acagccgtgt actactgcgc tagacactac 300 ggcgattact ggggccagggg aaccctggtc acagtgtcat ct 342 <210> 177 <211> 114 <212> PRT <213> Homo sapiens <400> 177 Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Arg Ser Asp Thr Ser As Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg His Tyr Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 100 105 110 Dear Dear <210> 178 <211> 342 <212> DNA <213> Homo sapiens <400> 178 cagctccagc tgcaagagtc tggacctgga ctggtcaagc ccagcgagac actgagcctg 60 acctgtaccg tgtccggcta cagcatcaca agcgactacg cctggaactg gatcagacag 120 cctcctggca aaggcctcga gtggatcggc tacatcagct acagcggcag aaccagctac 180 aaccccagcc tgaagtccag agtgaccatc agcagagaca ccagcaagaa ccagttctcc 240 ctgaagctga gcagcgtgac agccgccgat acagccgtgt actactgcgc tagacactac 300 ggcgattact ggggccaggg aaccctggtc acagtgtcat ct 342 <210> 179 <211> 114 <212> PRT <213> Homo sapiens <400> 179 Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 1 5 10 15 Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Thr Ser Asp 20 25 30 Tyr Ala Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp 35 40 45 Ile Gly Tyr Ile Ser Tyr Ser Gly Arg Thr Ser Tyr Asn Pro Ser Leu 50 55 60 Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser 65 70 75 80 Leu Lys Leu Ser Ser Val Thr Ala Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Only Arg His Tyr Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val 100 105 110 Ser Ser <210> 180 <211> 342 <212> DNA <213> Homo sapiens <400> 180 cagctccagc tgcaagagtc tggacctgga ctggtcaagc ccagcgagac actgagcctg 60 acctgtaccg tgtccggcta cagcatcaca agcgactacg cctggactg gatcagacag 120 cctcctggca aaggcctcga gtggatcggc tacatcagct acagcggcag aaccacctac 180 aaccccagcc tgaagtccag agtgaccacc agcgtagaca ccagcaagaa ccagttctcc 240 ctgaagctga gcagcgtgac agccgccgat acagccgtgt actactgcgc tagacactac 300 ggcgattact ggggccagggg aaccctggtc acagtgtcat ct 342 <210> 181 <211> 120 <212> PRT <213> Homo sapiens <400> 181 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Thr Tyr 20 25 30 Serving His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Trp Glu Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Thr Thr Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Tyr Tyr Thr Phe Tyr His Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Create Create 115 120 <210> 182 <211> 360 <212> DNA <213> Homo sapiens <400> 182 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcaagg ctagcggcta caccttcgcc acctacagca tccactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcaacccca gctctggcta caccaactac 180 aaccagaaat tccagggccg cgtgaccctc acagccgaca agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagagtgtac 300 tacacattct accacgccat ggactactgg ggccagggca caacagtgac cgttagctct 360 <210> 183 <211> 17 <212> PRT <213> Homo sapiens <400> 183 Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe Gln 1 5 10 15 Gly <210> 184 <211> 120 <212> PRT <213> Homo sapiens <400> 184 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Thr Tyr 20 25 30 Ser Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Leu Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Tyr Tyr Thr Phe Tyr His Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 185 <211> 360 <212> DNA <213> Homo sapiens <400> 185 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcagg ctagcggcta caccttcgcc acctacagca tccactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcacccca gctctggcta caccactac 180 aaccagaaat tccaggccg cgtgaccctc acagccgacg agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagagtgtac 300 tacacattct accacgccat ggactactgg ggccagggca cacagtgac cgttagctct 360 <210> 186 <211> 120 <212> PRT <213> Homo sapiens <400> 186 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Thr Tyr 20 25 30 Serving His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Trp Glu Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Only Asp Lys Sert Thr Thr On Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Tyr Tyr Thr Phe Tyr His Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 187 <211> 360 <212> DNA <213> Homo sapiens <400> 187 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcagg ctagcggcta caccttcgcc acctacagca tccactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcacccca gctctggcta caccactac 180 aaccagaaat tccaggccg cgtgaccatc acagccgaca agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagagtgtac 300 tacacattct accacgccat ggactactgg ggccagggca cacagtgac cgttagctct 360 <210> 188 <211> 120 <212> PRT <213> Homo sapiens <400> 188 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Thr Tyr 20 25 30 Ser Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Ser Gly Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Tyr Tyr Thr Phe Tyr His Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 189 <211> 360 <212> DNA <213> Homo sapiens <400> 189 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcagg ctagcggcta caccttcgcc acctacagca tccactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcacccca gctctggcta caccactac 180 aaccagaaat tccaggccg cgtgaccactac acagccgacg agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagagtgtac 300 tacacattct accacgccat ggactactgg ggccagggca cacagtgac cgttagctct 360 <210> 190 <211> 120 <212> PRT <213> Homo sapiens <400> 190 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg Ser His 20 25 30 Thr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Containing Pro Ser Thr Gly Tyr Containing Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Leu Thr Ala Asp Lys Ser Thr Thr Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Asn Tyr Ala Tyr Tyr Tyr Gly Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Create Create 115 120 <210> 191 <211> 360 <212> DNA <213> Homo sapiens <400> 191 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcaagg ctagcggcta caccttcaga tcccacacca tgcactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcaacccca gcaccggcta tatcaactac 180 aaccagaaat tccagggccg cgtgaccctc acagccgaca agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagaagcaac 300 tacgcctact actacggcat ggactactgg ggccagggca ccacagtgac agtctcttct 360 <210> 192 <211> 17 <212> PRT <213> Homo sapiens <400> 192 Tyr Ile Asn Pro Ser Thr Gly Tyr Ile Asn Tyr Asn Gln Lys Phe Gln 1 5 10 15 Gly <210> 193 <211> 120 <212> PRT <213> Homo sapiens <400> 193 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg Ser His 20 25 30 Thr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Thr Gly Tyr Ile Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Leu Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Asn Tyr Ala Tyr Tyr Tyr Gly Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 194 <211> 360 <212> DNA <213> Homo sapiens <400> 194 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcaagg ctagcggcta caccttcaga tcccacacca tgcactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcaacccca gcaccggcta tatcaactac 180 aaccagaaat tccagggccg cgtgaccctc acagccgacg agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagaagcaac 300 tacgcctact actacggcat ggactactgg ggccagggca ccacagtgac agtctcttct 360 <210> 195 <211> 120 <212> PRT <213> Homo sapiens <400> 195 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg Ser His 20 25 30 Thr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Containing Pro Ser Thr Gly Tyr Containing Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Only Asp Lys Sert Thr Thr On Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Only Arg Ser Asn Tyr Ala Tyr Tyr Gly Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 196 <211> 360 <212> DNA <213> Homo sapiens <400> 196 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcagg ctagcggcta caccttcaga tcccaccacca tgcactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcacccca gcaccggctta tatchactac 180 aaccagaaat tccaggccg cgtgaccatc acagccgaca agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagaagcac 300 tacgcctact actacggcat ggactactgg ggccagggca ccacagtgac agtctcttct 360 <210> 197 <211> 120 <212> PRT <213> Homo sapiens <400> 197 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Arg Ser His 20 25 30 Thr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Tyr Ile Asn Pro Ser Thr Gly Tyr Ile Asn Tyr Asn Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Asn Tyr Ala Tyr Tyr Tyr Gly Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <210> 198 <211> 360 <212> DNA <213> Homo sapiens <400> 198 caggttcagc tggttcagtc tggcgccgaa gtgaagaaac ctggcagcag cgtgaaggtg 60 tcctgcaagg ctagcggcta caccttcaga tcccacacca tgcactgggt ccgacaggct 120 ccaggacagg gacttgagtg gatgggctac atcaacccca gcaccggcta tatcaactac 180 aaccagaaat tccagggccg cgtgaccatc acagccgacg agtctacaag caccgcctac 240 atggaactga gcagcctgag aagcgaggac accgccgtgt actactgcgc cagaagcaac 300 tacgcctact actacggcat ggactactgg ggccagggca ccacagtgac agtctcttct 360 <210> 199 <211> 107 <212> PRT <213> Homo sapiens <400> 199 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 200 <211> 321 <212> DNA <213> Homo sapiens <400> 200 gataccaga tgacacag ccctagcagc ctgtctgcca gcgtgggaga cagagtgacc 60 atcacctgta gagccagcca gaacgtgggc acaaacgtgg cctggtatca gcagaagcct 120 ggcaaggctc ccaaggcgct gatctacagc gccagctaca gagagagcgg cgtgccatct 180 agattcagcg gctctggctc tgcaccgac ttcacctga caatctctag cctgcagcct 240 gaggacttcg ctacctactt ctgccagcag ttcaacaa acccttgac ctcggcgga 300 gghunter g 321 <210> 201 <211> 11 <212> PRT <213> Homo sapiens <400> 201 Arg Ala Ser Gln Asn Val Gly Thr Asn Val Ala 1 5 10 <210> 202 <211> 7 <212> PRT <213> Homo sapiens <400> 202 Ser Ala Ser Tyr Arg Glu Ser 1 5 <210> 203 <211> 107 <212> PRT <213> Homo sapiens <400> 203 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 204 <211> 321 <212> DNA <213> Homo sapiens <400> 204 gacatccaga tgacacagag ccctagcagc ctgtctgcca gcgtgggaga cagagtgacc 60 atcacctgta gagccagcca gaacgtgggc acaaacgtgg cctggtatca gcagaagcct 120 ggcaaggctc ccaagctgct gatctacagc gccagctaca gagagagcgg cgtgccatct 180 agattcagcg gctctggctc tggcaccgac ttcaccctga caatctctag cctgcagcct 240 gaggacttcg ctacctactt ctgccagcag ttcaacacaa accctctgac cttcggcgga 300 ggcaccaagg tggaaatcaa g 321 <210> 205 <211> 107 <212> PRT <213> Homo sapiens <400> 205 Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 206 <211> 321 <212> DNA <213> Homo sapiens <400> 206 gataccaga tgacacag ccctagcagc ctgtctgcca gcgtgggaga cagagtgacc 60 atcacctgta gagccagcca gaacgtgggc acaaacgtgg cctggtatca gcagaagcct 120 ggcaaggctc ccaaggcgct gatctacagc gccagctaca ggagagcgg cgtgccatct 180 agattcagcg gctctggctc tgcaccgac ttcacctga caatctctag cctgcagcct 240 gaggacttcg ctacctactt ctgccagcag ttcaacaa acccttgac ctcggcgga 300 ggcaccaagg tggaaatcaa g 321 <210> 207 <211> 7 <212> PRT <213> Homo sapiens <400> 207 Ser Ala Ser Tyr Arg Gln Ser 1 5 <210> 208 <211> 107 <212> PRT <213> Homo sapiens <400> 208 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 209 <211> 321 <212> DNA <213> Homo sapiens <400> 209 gagatcgtga tgacacagag ccccgccaca ctgagtgtgt ctccaggcga aagagctacc 60 ctgagctgta gagcctctca gaacgtgggc acaaacgtgg cctggtatca gcagaagcct 120 ggacaggctc ccagagcgct gatctacagc gccagctaca gagagtctgg catccctgct 180 agattcagcg gctctggctc tggcaccgag ttcaccctga caatcagcag cctgcagagc 240 gaggacttcg ccgtgtactt ctgccagcag ttcaacacaa accctctgac cttcggcgga 300 ggcaccaagg tggaaatcaa g 321 <210> 210 <211> 107 <212> PRT <213> Homo sapiens <400> 210 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Glu Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 211 <211> 321 <212> DNA <213> Homo sapiens <400> 211 gagatcgtga tgacacagag ccccgccaca ctgagtgtgt ctccaggcga aagagctacc 60 ctgagctgta gagccctca gaacgtgggc acaaacgtgg cctggtatca gcagaagcct 120 ggacaggctc ccagactgct gatctacagc gccagctaca gagagtctgg catccctgct 180 agattcagcg gctctggctc tggcaccgag ttcacctga caatcagcag cctgcagc 240 gaggacttcg ccgtgtactt ctgccagcag ttcaacaa acccttgac ctcggcgga 300 gghunter g 321 <210> 212 <211> 107 <212> PRT <213> Homo sapiens <400> 212 Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Asn Val Gly Thr Asn 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Ala Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Gln Ser Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80 Glu Asp Phe Ala Val Tyr Phe Cys Gln Gln Phe Asn Thr Asn Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 213 <211> 321 <212> DNA <213> Homo sapiens <400> 213 gagatcgtga tgacacagag ccccgccaca ctgagtgtgt ctccaggcga aagagctacc 60 ctgagctgta gagcctctca gaacgtgggc acaaacgtgg cctggtatca gcagaagcct 120 ggacaggctc ccagagcgct gatctacagc gccagctaca ggagctgg catccctgct 180 agattcagcg gctctggctc tggcaccgag ttcacctga caatcagcag cctgcagc 240 gaggacttcg ccgtgtactt ctgccagcag ttcaacaa acccttgac ctcggcgga 300 gghunter g 321 <210> 214 <211> 107 <212> PRT <213> Homo sapiens <400> 214 Asp Ile Gln Met Thr Gln Ser Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Is Thr Is Thr Cys Arg Is Only Gln Asp Is Only Thr 20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Tyr Thr Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln His Tyr Asn Ile Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 <210> 215 <211> 321 <212> DNA <213> Homo sapiens <400> 215 gacatccaga tgacacagag ccctagcagc ctgtctgcca gcgtgggaga cagagtgacc 60 atcacctgta gagccagcca ggacgtgaac acagccgtgg cttggtatca gcagaagcct 120 ggcaaggccc ctaagctgct gatctacagc gccagctaca gatacaccgg cgtgccctct 180 agattcagcg gctctggctc tggcaccgac ttcaccctga caatctctag cctgcagcct 240 gaggacttcg ccacctacta ctgccagcag cactacaaca tccctctgac cttcggcgga 300 ggcaccaagg tggaaatcaa g 321 <210> 216 <211> 11 <212> PRT <213> Homo sapiens <400> 216 Arg Ala Ser Gln Asp Val Asn Thr Ala Val Ala 1 5 10

Claims

1. An antibody or antigen-binding portion thereof that binds to MASP-2, comprising the heavy chain CDR1, CDR2 and CDR3 sequences shown in SEQ ID NOs: 7, 17 and 141, respectively, and the light chain CDR1, CDR2 and CDR3 sequences shown in SEQ ID NOs: 201, 202 and 146, respectively.

2. The antibody or antigen-binding portion thereof according to claim 1, comprising a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 177, and a light chain variable region having an amino acid sequence as shown in SEQ ID NO:

208.

3. A bispecific or multispecific molecule comprising the MASP-2 binding antibody of claim 1 or 2, or an antigen binding portion thereof.

4. The antibody or antigen binding portion thereof of claim 1 or 2 or the bispecific or multispecific molecule of claim 3, which is humanized.

5. A nucleic acid encoding the antibody or antigen binding portion thereof of claim 1 or 2, the bispecific or multispecific molecule of claim 3, or the antibody or antigen binding portion thereof or the bispecific or multispecific molecule of claim 4.

6. The nucleic acid according to claim 5, comprising a heavy chain variable region nucleotide sequence as shown in SEQ ID NO: 178, and a light chain variable region nucleotide sequence as shown in SEQ ID NO:

209.

7. A vector comprising the nucleic acid according to claim 5 or 6.

8. A cell comprising the nucleic acid of claim 5 or 6 or the vector of claim 7.

9. A composition comprising the antibody or antigen binding portion thereof of claim 1 or 2, the bispecific or multispecific molecule of claim 3, the antibody or antigen binding portion thereof or the bispecific or multispecific molecule of claim 4, the nucleic acid of claim 5 or 6, the vector of claim 7 and / or the cell of claim 8.

10. The composition according to claim 9, further comprising one or more additional therapeutic drugs; said additional drugs are selected from: anti-inflammatory and / or analgesic drugs, anti-restenotic drugs, other complement inhibitors, chondroprotective drugs and / or spasm inhibitors.

11. A kit comprising the antibody or antigen binding portion thereof of claim 1 or 2, the bispecific or multispecific molecule of claim 3, the antibody or antigen binding portion thereof or the bispecific or multispecific molecule of claim 4, the nucleic acid of claim 5 or 6, the vector of claim 7, the cell of claim 8 and / or the composition of claim 9 or 10.

12. Use of the antibody or antigen binding portion thereof of claim 1 or 2, the bispecific or multispecific molecule of claim 3, the antibody or antigen binding portion thereof or the bispecific or multispecific molecule of claim 4, the nucleic acid of claim 5 or 6, the vector of claim 7, the cell of claim 8 and / or the composition of claim 9 or 10 in the preparation of a medicament or kit for diagnosing, treating or preventing a disease associated with MASP-2-dependent complement activation.

13. The use of claim 12, wherein the disease associated with MASP-2-dependent complement activation is selected from the group consisting of MASP-2-dependent complement-mediated vascular conditions, ischemia-reperfusion injury, atherosclerosis, inflammatory gastrointestinal disorders, pulmonary conditions, extracorporeal reperfusion procedures, skeletal muscle conditions, renal conditions, skin conditions, organ or tissue transplantation, nervous system disorders or injuries, blood disorders, genitourinary tract conditions, diabetes, chemotherapy or radiation treatment, malignancy, endocrine disorders, coagulation disorders, or ophthalmic conditions; complement activation-related injuries and diseases caused by bacterial, fungal or viral infections.

14. The use according to claim 12, wherein the disease associated with MASP-2-dependent complement activation is an autoimmune inflammatory condition or an autoimmune disease.

15. The use according to claim 14, wherein the autoimmune inflammatory condition is selected from the group consisting of multiple sclerosis, type I diabetes, Hashimoto's thyroiditis, pernicious anemia, Addison's disease, myasthenia gravis, rheumatoid arthritis, uveitis, psoriasis, Guillain-Barre syndrome, Graves' disease, systemic lupus erythematosus, glomeronephritis, dermatomyositis, asthma, eczema, atopic dermatitis, contact dermatitis, other eczematous dermatitis, seborrheic dermatitis, rhinitis, lichen planus, pemplugus, bullous pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitis, erythema, eosinophilia, alopecia areata, arteriosclerosis, primary biliary cirrhosis and nephrotic syndrome.

16. The use according to claim 12, wherein the disease associated with MASP-2-dependent complement activation is selected from the group consisting of inflammatory bowel disease, coeliac disease, eosinophilic gastroenteritis, mastocytosis and food-related allergies.

17. The use according to claim 12, wherein the disease associated with MASP-2-dependent complement activation is selected from the group consisting of proctitis and inflammatory bowel disease.

18. The use according to claim 12, wherein the disease associated with MASP-2-dependent complement activation is selected from Chrohn's disease and ulcerative colitis.

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