Antibody binding specifically to ASM protein

Abstract

The present invention relates to an antibody specifically binding to the acid sphingomyelinase (ASM) protein and a use thereof. Particularly, the antibody or antigen-binding fragment thereof according to the present invention binds specifically to the ASM protein with high affinity, and thus can be used for detecting the ASM protein or diagnosing diseases arising from the overexpression of the ASM protein.

Description

Antibodies that specifically bind to the ASM protein

[0001] The present invention relates to an antibody that specifically binds to ASM (acid sphingomyelinase) protein.

[0002]

[0003] Sphingolipid metabolism regulates normal cell signaling, and abnormal changes in sphingolipid metabolism contribute to various neurodegenerative diseases, including Alzheimer's disease. Acid sphingomyelinase (ASM), an enzyme that regulates sphingolipid metabolism, is expressed in nearly all cell types and plays a crucial role in sphingolipid metabolism and cell membrane turnover.

[0004] In the brains of patients with neurodegenerative diseases such as Alzheimer's disease, the activity of the ASM protein is significantly increased compared to that of healthy individuals. In this regard, Korean Patent Registration No. 10-1521117 discloses that inhibiting the activity of overexpressed ASM protein or suppressing its expression can suppress β-amyloid accumulation and improve learning ability and memory, thereby treating neurodegenerative diseases. Furthermore, it has recently been discovered that ASM protein activity is also increased in neurological diseases such as depression, and suppressing the expression or activity of the ASM protein has the effect of alleviating depression.

[0005] However, no substances have been developed that directly inhibit the expression or activity of the ASM protein. However, several inhibitors that indirectly suppress ASM protein expression have been identified. For example, tricyclic antidepressants are used to treat depression, including amitriptyline, desipramine, and mipramine. Although these tricyclic antidepressants were not developed as ASM protein inhibitors, various studies have demonstrated that they exhibit ASM protein inhibitory effects. The main pharmacological mechanism of tricyclic antidepressants is to increase neurotransmitter activity by inhibiting the reuptake of neurotransmitters in neurons, and their action as ASM inhibitors has been identified as a secondary effect. However, tricyclic antidepressants can cause side effects such as blurred vision, increased photosensitivity, and vomiting due to their effects on the nervous system and neurons.

[0006]

[0007] An object of the present invention is to provide an antibody or an antigen-binding fragment thereof that specifically binds to an ASM protein.

[0008] Another object of the present invention is to provide a method for producing the antibody or antigen-binding fragment thereof.

[0009] Another object of the present invention is to provide a use of the antibody or antigen-binding fragment thereof for detecting ASM protein.

[0010]

[0011] To achieve the above purpose, the present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to ASM (acid sphingomyelinase) protein.

[0012] Additionally, the present invention provides a nucleic acid encoding the antibody or an antigen-binding fragment thereof.

[0013] In addition, the present invention provides an expression vector comprising the nucleic acid.

[0014] In addition, the present invention provides a host cell comprising the nucleic acid or expression vector.

[0015] In addition, the present invention provides a method for producing an antibody or an antigen-binding fragment thereof that specifically binds to an ASM protein, comprising a step of culturing the host cell to produce the antibody or an antigen-binding fragment thereof.

[0016] In addition, the present invention provides a composition and kit for detecting ASM protein comprising the antibody or an antigen-binding fragment thereof.

[0017] In addition, the present invention provides a method for detecting ASM protein, comprising a step of reacting the antibody or an antigen-binding fragment thereof with a sample.

[0018]

[0019] The antibody or antigen-binding fragment thereof according to the present invention can be used for detecting ASM protein or diagnosing a disease caused by overexpression of ASM protein by specifically binding to ASM protein with high binding affinity.

[0020]

[0021] Hereinafter, the present invention will be described in detail.

[0022] The present invention provides an antibody or an antigen-binding fragment thereof that specifically binds to ASM (acid sphingomyelinase) protein.

[0023] The term 'ASM (acid sphingomyelinase) protein' used herein refers to an enzyme belonging to the SMase (sphingomyelinase) family that regulates sphingolipid metabolism. The ASM protein catalyzes the process of decomposing sphingomyelin into ceramide and phosphorylcholine, and can be classified as alkaline, neutral, or acidic depending on the pH at which it exhibits optimal enzyme activity.

[0024] The ASM protein may include any type of ASM protein known in the art. Specifically, the ASM protein may be of mammalian origin, and more specifically, may be of human, monkey, rat, or mouse origin. Furthermore, the ASM protein may include any amino acid sequence known in the art as an ASM protein. For example, the ASM protein may be a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 139 or a nucleic acid encoding the same.

[0025] The above ASM protein may have one or more amino acids added, deleted, or substituted in the amino acid sequence set forth in SEQ ID NO: 139, as long as it maintains the same or corresponding biological activity. In this case, the amino acid substitution may be a conservative substitution performed within a range that does not or only slightly affects the charge, i.e., polarity or hydrophobicity, of the entire protein. In addition, the ASM protein may have 80% or more, 90% or more, 95% or more, 97% or more, or 99% or more homology with the amino acid sequence set forth in SEQ ID NO: 139.

[0026] The term 'antibody' as used herein refers to an immune protein that binds to an antigen and interferes with the action of the antigen or removes the antigen. The antibody may include all types of antibodies included in the general technical field. Specifically, the antibody may include IgM, IgD, IgG, IgA, and IgE, each of which may include a heavy chain produced from genes encoding heavy chain constant regions, namely μ, δ, γ, α, and ε. Typically, IgG is used in antibody technology, which is composed of isotypes such as IgG1, IgG2, IgG3, or IgG4, each of which may have different structural and functional properties. The antibody may include a humanized antibody comprising minimal sequence derived from a non-human antibody, a human antibody composed of sequence derived from a human, or a chimeric antibody in which sequences derived from different species are mixed.

[0027] The above IgG can form a very stable Y-shaped structure (about 150 kDa) made of two heavy chain proteins of about 50 kDa and two light chain proteins of about 25 kDa. The light and heavy chains that constitute the antibody can be divided into a variable region whose amino acid sequence is different among antibodies and a constant region whose amino acid sequence is the same. At this time, the heavy chain constant region includes CH1, hinge (H), CH2, and CH3 domains, and each domain is composed of two β-sheets and can be connected by an intramolecular disulfide bond. At this time, the two variable regions of the heavy chain and light chain are combined to form an antigen binding site, and one antigen binding site can exist on each of the two arms of the Y shape. In a full-length antibody, the part that can bind to an antigen is called Fab (antibody binding fragment), and the part that cannot bind to an antigen is called Fc (crystallizable fragment). Fab and Fc can be connected by a hinge.

[0028] In one embodiment of the present invention, the IgG may be human-derived IgG, and specifically may include a human IgG heavy chain constant region consisting of an amino acid sequence set forth in SEQ ID NO: 137 and a human light chain λ constant region consisting of an amino acid sequence set forth in SEQ ID NO: 138. In addition, the human-derived IgG may include a human IgG heavy chain constant region consisting of a base sequence set forth in SEQ ID NO: 140 and a human light chain λ constant region consisting of a base sequence set forth in SEQ ID NO: 141.

[0029] The antibody according to the present invention may include not only a full-length antibody but also an antigen-binding fragment thereof. Specifically, the antigen-binding fragment may refer to a portion excluding the Fc, which functions to transmit antigen-binding stimulation to cells, complement, etc. For example, the antigen-binding fragment of an antibody may include all of Fab, scFv, F(ab)2, and Fv, and may also include third-generation antibody fragments such as single-domain antibodies or minibodies.

[0030] For example, the antibody or antigen-binding fragment thereof may include a heavy chain variable region comprising a heavy chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 89, a heavy chain CDR2 comprising a polypeptide comprising an amino acid sequence set forth in SEQ ID NO: 90 in which five or fewer amino acids are substituted, and a heavy chain CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 91; and a light chain variable region comprising a light chain CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 92, a light chain CDR2 comprising a polypeptide comprising an amino acid sequence set forth in SEQ ID NO: 93 or in which five or fewer amino acids are substituted from the polypeptide, and a light chain CDR3 comprising a polypeptide comprising an amino acid sequence set forth in SEQ ID NO: 94 or in which two or fewer amino acids are substituted from the polypeptide.

[0031] The above term, 'CDR (complementarity determining region)' refers to a hypervariable region that has a different amino acid sequence for each antibody within the heavy and light chain variable regions of the antibody, and refers to the site that actually binds to the antigen. In the three-dimensional structure of the antibody, the CDR is located on the surface of the antibody in a loop shape, and a FR (framework region) that structurally supports it may exist below the loop. Both the heavy chain and the light chain have three loop structures each, and these six loop structures can come into direct contact with the antigen. The antigen-binding site CDR having the six loop structures may be conveniently referred to as heavy chain CDR1, heavy chain CDR2, heavy chain CDR3, light chain CDR1, light chain CDR2, or light chain CDR3, respectively.

[0032] In the antibody or antigen-binding fragment thereof according to the present invention, the heavy chain CDR2 in which five or fewer amino acids are substituted in a polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 90 may be a polypeptide in which five or fewer amino acids are substituted selected from the group consisting of the 2nd, 4th to 10th, 12th, 13th, and 17th amino acids from the N-terminus of the polypeptide.

[0033] For example, the 2nd, 4th, 6th, 7th, 9th, 12th or 13th amino acid from the N-terminus of the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 90 may be substituted with a neutral amino acid, and specifically, the neutral amino acid may include alanine, glycine, leucine, isoleucine, proline, valine, phenylalanine, tryptophan, tyrosine, serine, threonine, cysteine, methionine, asparagine and glutamine. More specifically, the second amino acid from the N-terminus, isoleucine, may be substituted with leucine, valine, or threonine, the fourth amino acid, tyrosine, may be substituted with phenylalanine or tryptophan, the sixth amino acid, serine, may be substituted with glycine or asparagine, the seventh amino acid, glycine, may be substituted with valine, the ninth amino acid, isoleucine, may be substituted with proline, alanine, or valine, the twelfth amino acid, alanine, may be substituted with serine, and the thirteenth amino acid, aspartic acid, may be substituted with asparagine or alanine. Meanwhile, the 10th amino acid from the N-terminus of the polypeptide composed of the amino acid sequence described in SEQ ID NO: 90 may be substituted with a basic amino acid, and specifically, the basic amino acid may include arginine, histidine, and lysine. More specifically, tyrosine, which is the 10th amino acid from the N-terminus, may be substituted with arginine.In addition, the 17th amino acid from the N-terminus of the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 90 may be substituted with an acidic amino acid, and specifically, the acidic amino acid may include aspartic acid and glutamic acid. More specifically, glycine, which is the 17th amino acid from the N-terminus, may be substituted with aspartic acid or glutamic acid. In addition, the 5th amino acid from the N-terminus of the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 90 may be a neutral or acidic amino acid, and the neutral or acidic amino acid may be as described above. More specifically, glycine, which is the 5th amino acid from the N-terminus, may be substituted with alanine, aspartic acid, serine, or proline. Furthermore, the eighth amino acid from the N-terminus of the polypeptide comprising the amino acid sequence described in SEQ ID NO: 90 may be a neutral, acidic or basic amino acid, and the neutral, acidic or basic amino acid may be as described above. More specifically, asparagine, which is the eighth amino acid from the N-terminus, may be substituted with aspartic acid, lysine, isoleucine, threonine, valine, glycine or tyrosine.

[0034] In one embodiment of the present invention, the heavy chain CDR2 in which five or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 90 may be a polypeptide consisting of the amino acid sequences described in SEQ ID NOs: 95 to 111, respectively.

[0035] In the antibody or antigen-binding fragment thereof according to the present invention, the light chain CDR2 in which five or fewer amino acids are substituted in a polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 93 may be a polypeptide in which four or fewer amino acids selected from the group consisting of amino acids 3 to 7 from the N-terminus of the polypeptide are substituted.

[0036] For example, the 3rd, 4th, and 5th amino acids from the N-terminus of the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 93 may be neutral amino acids, and the neutral amino acids may be as described above. More specifically, the 3rd amino acid from the N-terminus, serine, may be substituted with isoleucine, leucine, methionine, glutamine, threonine, valine, tryptophan, or tyrosine, the 4th amino acid, histidine, may be substituted with phenylalanine or tyrosine, and the 5th amino acid, arginine, may be substituted with glutamine, proline, leucine, threonine, or serine. Meanwhile, the 6th amino acid from the N-terminus of the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 93 may be a basic or neutral amino acid, and the basic or neutral amino acids may be as described above. More specifically, the 6th amino acid from the N-terminus, proline, may be substituted with alanine, glutamine, arginine, serine, or histidine. Furthermore, the 7th amino acid from the N-terminus of the polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 93 may be a neutral, acidic, or basic amino acid, and the neutral, acidic, or basic amino acid may be as described above. More specifically, the 7th amino acid from the N-terminus, serine, may be substituted with aspartic acid, tyrosine, glycine, asparagine, threonine, phenylalanine, tryptophan, or arginine.

[0037] In one embodiment of the present invention, the light chain CDR2 in which five or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 93 may be a polypeptide consisting of the amino acid sequences described in SEQ ID NOs: 112 to 135, respectively.

[0038] In the antibody or antigen-binding fragment thereof according to the present invention, the light chain CDR2 in which two or fewer amino acids are substituted in a polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 94 may be a polypeptide in which two or fewer amino acids are substituted selected from the group consisting of the 6th and 8th amino acids from the N-terminus of the polypeptide.

[0039] For example, the 6th and 8th amino acids from the N-terminus of the polypeptide comprising the amino acid sequence described in SEQ ID NO: 94 may be neutral amino acids, and the neutral amino acids may be as described above. More specifically, serine, which is the 6th and 8th amino acid from the N-terminus, may be substituted with tryptophan and glycine, respectively.

[0040] In one embodiment of the present invention, the light chain CDR3 in which two or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 94 may be a polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 136.

[0041] In addition, the antibody or antigen-binding fragment thereof according to the present invention may be modified as needed. Specifically, the antibody or antigen-binding fragment thereof may be modified by conjugation, glycosylation, labeling, or a combination thereof. Specifically, the antibody or antigen-binding fragment thereof may be modified with horseradish peroxidase (HRP), alkaline phosphatase, hapten, biotin, streptavidin, fluorescent substances, radioactive substances, quantum dots, polyethylene glycol (PEG), histidine labeling, etc. In addition, the antibody or antigen-binding fragment thereof may be conjugated with other drugs as needed.

[0042] The above antibody or antigen-binding fragment thereof can be produced according to a monoclonal antibody production method known in the art, and the production method can be appropriately modified by those skilled in the art. For example, the antibody can be produced by producing a hybridoma using B lymphocytes obtained from an animal immunized with an antigen, or produced using phage display technology.

[0043]

[0044] Additionally, the present invention provides a nucleic acid encoding the antibody or an antigen-binding fragment thereof.

[0045] The antibody or antigen-binding fragment thereof encoded by the nucleic acid according to the present invention may have the characteristics described above. As long as the amino acid sequence constituting the antibody or antigen-binding fragment thereof according to the present invention is known, the nucleic acid sequence encoding it will also be apparent to those skilled in the art. Furthermore, the nucleic acid sequence may have one or more bases added, deleted, or substituted, as long as the activity of the antibody or antigen-binding fragment thereof translated therefrom is maintained.

[0046]

[0047] In addition, the present invention provides an expression vector comprising the nucleic acid.

[0048] The nucleic acid included in the expression vector according to the present invention may encode an antibody or an antigen-binding fragment thereof having the characteristics described above.

[0049] The term 'expression vector' used herein refers to a means for expressing a target gene in a host cell, and may include plasmid vectors, cosmid vectors, bacteriophage vectors, viral vectors, and the like. The expression vector may include elements necessary for producing a peptide from the nucleic acid contained therein. Specifically, the expression vector may include a signal sequence, a replication origin, a marker gene, a promoter, a transcription termination sequence, and the like. In this case, a nucleic acid encoding an antibody or an antigen-binding fragment thereof according to the present invention may be operably linked to a promoter.

[0050] For example, an expression vector used in prokaryotic cells may include a promoter to initiate transcription, a ribosome binding site to initiate translation, and a transcription and translation termination sequence. Meanwhile, an expression vector used in eukaryotic cells may include a promoter and polyadenylation sequence derived from mammalian or mammalian viruses.

[0051] In addition, any marker gene known in the art may be used as the marker gene included in the expression vector, and specifically, it may be an antibiotic resistance gene. Specifically, the antibiotic resistance gene may be a gene that exhibits resistance to antibiotics including ampicillin, gentamicin, carbenicillin, chloramphenicol, streptomycin, kanamycin, neomycin, tetracycline, and the like.

[0052]

[0053] In addition, the present invention provides a host cell comprising the nucleic acid or expression vector.

[0054] The nucleic acid or expression vector included in the host cell according to the present invention may have the characteristics described above. For example, the nucleic acid may encode an antibody or an antigen-binding fragment thereof that specifically binds to the ASM protein according to the present invention, and the expression vector may include the nucleic acid described above.

[0055] The host cell may be any type of cell known to be capable of producing antibodies or antigen-binding fragments thereof in the art. Specifically, the host cell may be a prokaryotic cell, yeast, or eukaryotic cell. The prokaryotic cell may include Escherichia coli (E. coli), a strain of the genus Bacillus, a strain of the genus Streptomyces, a strain of the genus Pseudomonas, a strain of the genus Staphylococcus, and the like, and the yeast may include Saccharomyces cerevisiae, and the like. Meanwhile, the eukaryotic cells may include COS-7, BHK, CHO, CHOK1, DXB-11, DG-44, CHO / -DHFR, CV1, HEK293, TM4, VERO, HELA, MDCK, BRL 3A, W138, Hep G2, SK-Hep, MMT, TRI, MRC 5, FS4, 3T3, RIN, A549, PC12, K562, PERC6, SP2 / 0, NS-0, U20S, and HT1080.

[0056] In addition, the host cell may be transfected with the nucleic acid or expression vector described above according to a method known in the art. Specifically, the transfection may be performed by a method such as transient transfection, microinjection, transduction, cell fusion, calcium phosphate precipitation, liposome-mediated transfection, DEAE dextran-mediated transfection, polybrene-mediated transfection, electroporation, or a gene gun. In addition, the method may be appropriately modified by a person skilled in the art.

[0057]

[0058] In addition, the present invention provides a method for producing an antibody or an antigen-binding fragment thereof that specifically binds to an ASM protein, comprising a step of culturing the host cell to produce the antibody or an antigen-binding fragment thereof.

[0059] An antibody or antigen-binding fragment thereof produced by the production method according to the present invention may have the characteristics described above.

[0060] The above cultivation can be performed using an appropriate culture medium depending on the type of host cell used for production, and may include appropriate supplements as needed. Furthermore, the above cultivation can be performed in an appropriate environment depending on the type of host cell.

[0061] The production method according to the present invention may further include a step of recovering an antibody or antigen-binding fragment thereof produced in a host cell. This recovery may be performed using methods known in the art, which may be appropriately modified by those skilled in the art as needed. For example, this recovery may be performed by removing impurities using centrifugation or ultrafiltration, and further purifying the resulting product using chromatography or the like. The chromatography may include affinity chromatography, cation chromatography, hydrophobic interaction chromatography, or the like.

[0062]

[0063] In addition, the present invention provides a composition and kit for detecting ASM protein comprising the antibody or an antigen-binding fragment thereof.

[0064] The antibody or antigen-binding fragment thereof included in the composition and kit for detecting ASM protein according to the present invention may have the characteristics described above.

[0065] Additionally, the composition may include a ligand capable of specifically binding to the antibody or antigen-binding fragment thereof according to the present invention. The ligand may be a conjugate labeled with a detectable substance such as a chromogenic enzyme, a fluorescent substance, a radioisotope, or a colloid, or a ligand treated with streptavidin or avidin. In addition to the reagents described above, the detection composition of the present invention may further include distilled water or a buffer that stably maintains the structure of the reagents.

[0066] Additionally, the kit may be bound to a solid substrate to facilitate subsequent steps, such as washing the antibody or antigen-binding fragment thereof contained therein or separating the complex. The solid substrate may include, but is not limited to, a synthetic resin, nitrocellulose, a glass substrate, a metal substrate, glass fiber, microspheres, or microbeads. Furthermore, the synthetic resin may include polyester, polyvinyl chloride, polystyrene, polypropylene, PVDF, or nylon.

[0067] Additionally, the kit can be manufactured by a conventional manufacturing method known to those skilled in the art, and may further include a buffer, a stabilizer, an inactive protein, etc.

[0068]

[0069] In addition, the present invention provides a method for detecting ASM protein, comprising a step of reacting the antibody or an antigen-binding fragment thereof with a sample.

[0070] The antibody or antigen-binding fragment thereof used in the ASM protein detection method according to the present invention may have the characteristics described above.

[0071] The above sample may include any type of sample for which the ASM protein is to be detected. Furthermore, methods for detecting target proteins using antibodies or antigen-binding fragments thereof are well known in the art, and can be appropriately modified and implemented by those skilled in the art as needed.

[0072]

[0073] Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended only to illustrate the present invention and are not intended to limit the present invention. Anything that has substantially the same structure as the technical idea described in the claims of the present invention and achieves the same operational effects is included within the technical scope of the present invention.

[0074]

[0075] Example 1. Production of an antibody that specifically binds to ASM (acid sphingomyelinase) protein

[0076] A mutant was prepared from antibody #9104, an antibody that specifically binds to human ASM protein (SEQ ID NO: 139).

[0077] First, the VH and VL genes of the #9104 antibody were amplified by a conventional method using random primers to induce random mutations in the heavy and light chain CDR sequences of the #9104 antibody. The amplified VH and VL genes were each linked to the mouse heavy chain constant region 1 (CH1, SEQ ID NO: 142) and light chain constant region (CL, SEQ ID NO: 143) and inserted into the pComb3xss phagemid vector in the form of scFab. Using this, a #9104 random mutagenesis library was constructed, and screening was performed using the library by a conventional method to select scFabs that specifically bind to human ASM protein. An expression vector was constructed so that the selected scFab was expressed in a form in which a human heavy chain constant region and a human light chain λ constant region, each consisting of the base sequences described in SEQ ID NO: 137 or 138, were linked to the carboxy terminus of the heavy chain variable region and the light chain variable region, respectively. As a result, the amino acid sequence and nucleic acid sequence of the heavy chain variable region of the selected scFab are shown in Table 1 below, and the amino acid sequence and nucleic acid sequence of the light chain variable region are shown in Table 2 below.

[0078] 항체서열서열번호#9104EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGSGNIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 1GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTCTCCAACTACTACATGTCCTGGGTCCGACAGGCCCCTGGCAAAGGATTGGAATGGGTGTCCGGCATCTACTACGGCTCCGGCAACATCTACTATGCCGACTCCGTGAAGGGCAGATTCACCATCTCTCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGAGAGCCGAGGACACCGCCGTGTACTACTGCGCTAGAGATACCCCTGGCTTCGACTATTGGGGCCAGGGCACACTGGTCACCGTGAGCTCA서열번호 2#9104v-H02EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYFGSGDIRYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 3GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGAGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTATTTTGGTAGTGGTGATATAAGGTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호4#9104v-H05EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGGVKIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 5GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTATTATGGTGGTGTTAAAATATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 6#9104v-H10EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGNGIPYYADSVKDRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 7GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGTATCTATTATGGTAATGGTATTCCATATTACGCTGATTCTGTAAAAGATCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호8#9104v-H11EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGSGIPYYADSVKERFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 9GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTATTATGGTAGTGGTATTCCATATTACGCTGATTCTGTAAAAGAACGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 10#9104v-H13EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGSGIPYYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 11GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTATTATGGTAGTGGTATACCCTATTACGCTAATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호12#9104v-H14EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGSGKPYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 13GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTATTATGGTAGTGGTAAACCATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 14#9104v-H17EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRRAPGKGLEWVSGIYYGSGTPYYAASVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 15GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCGGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTATTATGGTAGTGGTACTCCATATTACGCTGCTTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호16#9104v-H18EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGSGTPYYSDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 17GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTACTATGGTAGTGGTACTCCATACTACTCTGACTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 18#9104v-H19EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGIYYGSGVPYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 19GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGATCTACTATGGCAGTGGTGTTCCATATTACGCTGATTCTGTCAAAGGGCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호20#9104v-H22EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGLYYGGGTPYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 21GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGCTCTATTATGGTGGTGGTACTCCATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 22#9104v-H24EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGLYYGSGIPYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 23GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGTTGTATTATGGTAGTGGTATTCCATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호24#9104v-H25EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGTYYASGGIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 25GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGACATATTATGCTAGTGGTGGTATATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 26#9104v-H26EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGTYYASGNAYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 27GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGACCTATTATGCTAGTGGTAATGCATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호28#9104v-H27EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGTYYDSGYVYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 29GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGGTTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGACCTATTATGATAGTGGATATGTATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 30#9104v-H30EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGTYYSSGGIYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 31GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGACCTATTATAGTAGTGGTGGTATATATTACGCTGATTCTGTCAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호32#9104v-H31EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGVYYGSGTPYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 33GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGGTCTACTATGGTAGTGGTACTCCATATTACGCTGATTCTGTAAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 34#9104v-H33EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVSGVYYPSGNPYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDTPGFDYWGQGTLVTVSS서열번호 35GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAATTATTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGGGTATATTATCCTAGTGGTAATCCATATTACGCTGATTCTGTTAAAGGTCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGATACGCCTGGGTTCGACTACTGGGGCCAGGGTACACTGGTCACCGTGAGCTCA서열번호 36

[0079] 항체서열서열번호#9104QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSHRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 37CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGCACACCTGGCCAGAGAGTGACCATCTCTTGTACCGGCTCCTCCTCCAACATCGGCAACAACGCCGTGAACTGGTATCAGCAGCTGCCTGGCACAGCCCCTAAACTGCTGATCTACTACGACTCCCACCGGCCTAGCGGCGTGCCCGATAGATTTTCCGGCTCTAAGTCCGGCACCTCTGCCAGCCTGGCTATCTCTGGACTGAGATCTGAGGACGAGGCCGACTACTACTGTGGCGCCTGGGACTACTCTCTGTCCGCCTATGTTTTTGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 38#9104v-L11QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDIFQPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 39CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATATTTTTCAGCCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호40#9104v-L12QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDIFRPDGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 41CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATATTTTTCGGCCAGACGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 42#9104v-L15QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDIHRPYGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 43CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATATTCATCGGCCATACGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호44#9104v-L19QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDLFRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 45CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATCTTTTTAGGCCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 46#9104v-L20QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDMFRPGGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 47CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTACGATATGTTCCGGCCAGGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호48#9104v-L21QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDMFRPNGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 49CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATATGTTCCGGCCAAACGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 50#9104v-L23QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDQFRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 51CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATCAATTTCGGCCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호52#9104v-L25QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSFPASGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 53CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTTTCCGGCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 54#9104v-L27QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSFRQSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 55CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTTTCGGCAAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호56#9104v-L28QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSFRRSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 57CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTTTCGGCGAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 58#9104v-L29QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSFRSTGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 59CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTACGATAGTTTTCGGTCAACCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호60#9104v-L32QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYLPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 61CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATCTGCCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 62#9104v-L33QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYPPYGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 63CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATCCGCCATACGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호64#9104v-L34QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYQASGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 65CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATCAGGCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 66#9104v-L37QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYRASGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 67CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATCGGGCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호68#9104v-L38QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYRPFGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 69CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATCGGCCATTCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 70#9104v-L39QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYRPGGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 71CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGACAGTTATCGGCCAGGGGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호72#9104v-L46QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYRSWGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 73CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATCGGTCATGGGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 74#9104v-L47QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSYTPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 75CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTTATACGCCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호76#9104v-L49QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDTFRPTGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 77CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATACTTTCCGGCCAACCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 78#9104v-L52QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDVHRPRGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 79CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATGTTCATCGGCCCAGGGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호80#9104v-L55QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDWYRPTGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 81CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATTGGTATCGGCCAACCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 82#9104v-L63QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDSHRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYWLGAYVFGGGTKLTVL서열번호 83CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATAGTCATCGGCCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATTAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATTGGCTGGGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호84#9104v-L64QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDYFSHSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 85CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATTATTTTAGCCATAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 86#9104v-L65QSVLTQPPSASGTPGQRVTISCTGSSSNIGNNAVNWYQQLPGTAPKLLIYYDYYQSSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCGAWDYSLSAYVFGGGTKLTVL서열번호 87CAGTCTGTGCTGACTCAGCCACCCTCAGCTAGCGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTACTGGCTCTTCATCTAATATTGGCAATAATGCTGTCAACTGGTACCAGCAGCTCCCAGGAACGGCCCCCAAACTCCTCATCTATTATGATTATTATCAGTCAAGCGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGTCCGAGGATGAGGCTGATTATTACTGTGGTGCTTGGGATTATAGCCTGAGTGCTTATGTCTTCGGCGGAGGCACCAAGCTTACGGTCCTA서열번호 88

[0080]

[0081] At this time, a mammalian expression vector was used as the expression vector. The produced expression vector was transformed into an ExpiCHO cell line to produce a full-length antibody that binds to human ASM protein.

[0082]

[0083] Example 2. Determination of complementarity determining region (CDR)

[0084] The complementarity determining region in the scFv manufactured above was confirmed by a conventional method, and as a result, the CDR sequence of the heavy chain variable region is shown in Table 3, and the CDR sequence of the light chain variable region is shown in Table 4.

[0085] Antibody CDR sequence SEQ ID NO: #9104CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGNIYYADSVKGSEQ ID NO: 90CDR3DTPGFDYSEQ ID NO: 91#9104v-H02CDR1NYYMSSEQ ID NO: 89CDR2GIYFGSGDIRYADSVKGSEQ ID NO: 95CDR3DTPGFDYSEQ ID NO: 91#9104v-H05CDR1NYYMSSEQ ID NO: 89CDR2GIYYGGVKIYYADSVKGSEQ ID NO: 96CDR3DTPGFDYSEQ ID NO: 91#9104v-H10CDR1NYYMSSEQ ID NO: 89CDR2GIYYGNGIPYYADSVKDSEQ ID NO: 97CDR3DTPGFDYSEQ ID NO: 91#9104v-H11CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGIPYYADSVKESEQ ID NO: 98CDR3DTPGFDYSEQ ID NO: 91#9104v-H13CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGIPYYANSVKGSEQ ID NO: 99CDR3DTPGFDYSEQ ID NO: 91#9104v-H14CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGKPYYADSVKGSEQ ID NO: 100CDR3DTPGFDYSEQ ID NO: 91#9104v-H17CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGTPYYAASVKGSEQ ID NO: 101CDR3DTPGFDYSEQ ID NO: 91#9104v-H18CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGTPYYSDSVKGSEQ ID NO: 102CDR3DTPGFDYSEQ ID NO: 91#9104v-H19CDR1NYYMSSEQ ID NO: 89CDR2GIYYGSGVPYYADSVKGSEQ ID NO: 103CDR3DTPGFDYSEQ ID NO: 91#9104v-H22CDR1NYYMSSEQ ID NO: 89CDR2GLYYGGGTPYYADSVKGSEQ ID NO: 104CDR3DTPGFDYSEQ ID NO: 91#9104v-H24CDR1NYYMSSEQ ID NO: 89CDR2GLYYGSGIPYYADSVKGSEQ ID NO: 105CDR3DTPGFDYSEQ ID NO: 91#9104v-H25CDR1NYYMSSEQ ID NO: 89CDR2GTYYASGGIYYADSVKG SEQ ID NO: 106CDR3DTPGFDY SEQ ID NO: 91#9104v-H26CDR1NYYMS SEQ ID NO: 89CDR2GTYYASGNAYYADSVKG SEQ ID NO:107CDR3DTPGFDYSEQ ID NO: 91#9104v-H27CDR1NYYMSSEQ ID NO: 89CDR2GTYYDSGYVYYADSVKGSEQ ID NO: 108CDR3DTPGFDYSEQ ID NO: 91#9104v-H30CDR1NYYMSSEQ ID NO: 89CDR2GTYYSSGGIYYADSVKGSEQ ID NO: 109CDR3DTPGFDYSEQ ID NO: 91#9104v-H31CDR1NYYMSSEQ ID NO: 89CDR2GVYYGSGTPYYADSVKGSEQ ID NO: 110CDR3DTPGFDYSEQ ID NO: 91#9104v-H33CDR1NYYMSSEQ ID NO: 89CDR2GVYYPSGNPYYADSVKGSEQ ID NO: 111CDR3DTPGFDY SEQ ID NO: 91

[0086] Antibody CDR sequence SEQ ID NO: 9104CDR1TGSSSNIGNNAVN SEQ ID NO: 92CDR2YDSHRPS SEQ ID NO: 93CDR3GAWDYSLSAYV SEQ ID NO: 94#9104v-L11CDR1TGSSSNIGNNAVN SEQ ID NO: 92CDR2YDIFQPS SEQ ID NO: 112CDR3GAWDYSLSAYV SEQ ID NO: 94#9104v-L12CDR1TGSSSNIGNNAVN SEQ ID NO: 92CDR2YDIFRPD SEQ ID NO: 113CDR3GAWDYSLSAYV SEQ ID NO: 94#9104v-L15CDR1TGSSSNIGNNAVN SEQ ID NO: 92CDR2YDIHRPY SEQ ID NO: 114CDR3GAWDYSLSAYV SEQ ID NO: 94#9104v-L19CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDLFRPSSEQ ID NO: 115CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L20CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDMFRPGSEQ ID NO: 116CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L21CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDMFRPNSEQ ID NO: 117CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L23CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDQFRPSSEQ ID NO: 118CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L25CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSFPASSEQ ID NO: 119CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L27CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSFRQSSEQ ID NO: 120CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L28CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSFRRSSEQ ID NO: 121CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L29CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSFRSTSEQ ID NO: 122CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L32CDR1TGSSSNIGNNAVN SEQ ID NO: 92CDR2YDSYLPS SEQ ID NO: 123CDR3GAWDYSLSAYV SEQ ID NO:94#9104v-L33CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYPPYSEQ ID NO: 124CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L34CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYQASSEQ ID NO: 125CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L37CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYRASSEQ ID NO: 126CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L38CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYRPFSEQ ID NO: 127CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L39CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYRPGSEQ ID NO: 128CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L46CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYRSWSEQ ID NO: 129CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L47CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSYTPSSEQ ID NO: 130CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L49CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDTFRPTSEQ ID NO: 131CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L52CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDVHRPRSEQ ID NO: 132CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L55CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDWYRPTSEQ ID NO: 133CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L63CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDSHRPSSEQ ID NO: 93CDR3GAWDYWLGAYVSEQ ID NO: 136#9104v-L64CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDYFSHSSEQ ID NO: 134CDR3GAWDYSLSAYVSEQ ID NO: 94#9104v-L65CDR1TGSSSNIGNNAVNSEQ ID NO: 92CDR2YDYYQSSSEQ ID NO: 135CDR3GAWDYSLSAYVSEQ ID NO: 94

[0087]

[0088] As shown in Tables 3 and 4, it was confirmed that CDR1 and CDR3 of the heavy chain variable region and CDR1 of the light chain variable region have the same sequence as the #9104 antibody, whereas CDR2 of the heavy chain variable region and CDR2 and CDR3 of the light chain variable region have sequences in which some of the amino acids constituting them are substituted.

[0089]

[0090] Experimental Example 1. Confirmation of binding affinity with ASM protein

[0091] The binding affinity and interaction kinetics of the antibody specifically binding to the ASM protein prepared above were determined by Octet ® Measurements were made using the QK384 system (Pall Life Sciences).

[0092] First, an antibody specifically binding to the ASM protein prepared in Example 1 was captured using an anti-human IgG Fc capture (AHC) biosensor, and 1.25, 2.5, 5, 10, or 20 nM of a recombinant human ASM protein solution was added thereto. After adding the human ASM protein solution, the binding phase of the reaction product was observed for about 1,200 seconds, and then 1× kinetics buffer (ForteBio) was added, and the separation phase of the reaction product was observed for about 1,500 seconds. Octet ® The association constant (K) for each antibody was determined using analysis software (Pall Life Sciences). a ), dissociation constant (K d ) and equilibrium dissociation constant (K D) was determined. As a result, the binding affinity of the antibody with the heavy chain CDR portion described in Table 1 mutated in the #9104 antibody to human ASM protein is shown in Table 5, and the binding affinity of the antibody with the light chain CDR portion described in Table 2 mutated to human ASM protein is shown in Table 6.

[0093] Antibody K D (M)K a (1 / Ms)K d (1 / s)#91042.25E-093.89E+058.73E-04#9104v-H021.20E-092.78E+053.33E-04#9104v-H 051.03E-091.71E+051.77E-04#9104v-H106.51E-102.37E+051.54E-04#9104v-H117.81E-1 02.50E+051.95E-04#9104v-H138.03E-102.31E+051.85E-04#9104v-H148.57E-103.28E+0 52.81E-04#9104v-H177.19E-103.38E+052.43E-04#9104v-H182.69E-099.79E+042.64E-04 #9104v-H196.92E-102.66E+051.84E-04#9104v-H227.98E-101.97E+051.57E-04#9104v-H 241.01E-092.04E+052.06E-04#9104v-H259.17E-102.07E+051.90E-04#9104v-H261.74E-0 91.75E+053.05E-04#9104v-H275.98E-102.76E+051.65E-04#9104v-H307.62E-101.99E+0 51.51E-04#9104v-H315.32E-102.66E+051.41E-04#9104v-H335.17E-102.08E+051.08E-04

[0094] Antibody K D (M)K a (1 / Ms)K d(1 / s)#91042.25E-093.89E+058.73E-04#9104v-L117.68E-102.59E+051.99E-04#9104v-L127.18E-102.21E+051.59E-04#9104v-L 159.92E-102.75E+052.73E-04#9104v-L191.03E-092.31E+052.38E-04#9104v-L206.58E-102.47E+051.62E-04#9104v-L215.56E-1 02.43E+051.35E-04#9104v-L238.17E-102.26E+051.85E-04#9104v-L258.82E-102.94E+052.59E-04#9104v-L271.02E-093.48E+0 53.55E-04#9104v-L281.05E-092.41E+052.54E-04#9104v-L291.07E-092.72E+052.92E-04#9104v-L321.37E-093.21E+054.39E-04 #9104v-L331.27E-092.27E+052.89E-04#9104v-L341.61E-093.06E+054.93E-04#9104v-L371.27E-092.59E+053.27E-04#9104v-L 381.80E-091.91E+053.43E-04#9104v-L391.52E-092.91E+054.43E-04#9104v-L468.80E-102.93E+052.58E-04#9104v-L471.41E-0 92.94E+054.15E-04#9104v-L497.85E-102.86E+052.25E-04#9104v-L529.82E-102.69E+052.64E-04#9104v-L558.79E-101.93E+0 51.70E-04#9104v-L631.41E-091.81E+052.55E-04#9104v-L645.05E-103.83E+051.93E-04#9104v-L655.32E-103.60E+051.92E-04

[0095]

[0096] As shown in Tables 5 and 6, the antibodies prepared in Example 1 were 10 -10 10 inland-9 It bound to human ASM protein with a binding affinity of M level.

[0097]

[0098] Experimental Example 2. Combination of heavy and light chain variable regions

[0099] Forty antibodies were produced by combining eight heavy chain variable regions and five light chain variable regions of the #9104 antibody selected above (Table 7).

[0100] Antibodies 가변영역#9104a-A1#9104v-H10#9104v-L20#9104a-A2#9104v-H11#9104a-A3#9104v-H13#9104a-A4#9104v-H17#9104a-A5#9104a-A5 104v-H19#9104a-A6#9104v-H27#9104a-A7#9104v-H31#9104a-A8#9104v-H33#9104a-B1#9104v-H10#9104v-L21#9104a-B 2#9104v-H11#9104a-B3#9104v-H13#9104a-B4#9104v-H17#9104a-B5#9104v-H19#9104a-B6#9104v-H27#9104a-B7#9104v-H31#9104a-B8#9104v-H33#9104a-C1#9104v-H10#9104v-L23#9104a-C2#9104v-H11#9104a-C3#9104v-H13#9104a-C4#91 04v-H17#9104a-C5#9104v-H19#9104a-C6#9104v-H27#9104a-C7#9104v-H31#9104a-C8#9104v-H33#9104a-D1#9104v-H10#9104v-L64#9104a-D2#9104v-H11#9104a-D3#9104v-H13#9104a-D4#9104v-H17#9104a-D5#9104v-H19#9104a-D6#9104v -H27#9104a-D7#9104v-H31#9104a-D8#9104v-H33#9104a-E1#9104v-H10#9104v-L65#9104a-E2#9104v-H11#9104a-E3#9104v-H13#9104a-E4#9104v-H17#9104a-E5#9104v-H19#9104a-E6#9104v-H27#9104a-E7#9104v-H31#9104a-E8#9104v-H33

[0101]

[0102] As described above, after small-scale expression in ExpiCHO cell lines, the binding affinity to human ASM protein was confirmed as described in Experimental Example 1. At this time, the antibody prepared by combining the #9104v-H33 heavy chain variable region and the #9104v-L23 light chain variable region was not expressed and was therefore excluded from the binding affinity analysis with ASM protein. As a result, the binding affinity of the antibody to human ASM protein is shown in Table 8.

[0103] Antibody K D (M)K a (1 / Ms)K d(1 / s)#91042.19E-093.91E+058.55E-04#9104a-A15.18E-112.20E+051.14E-05#9104a-A29.61E-112.06E+051.98E-05#9104a-A32.77E-111.98E+055.48E-06#9104a-A41.62E-102.19E+053.54E-05#9104a-A51.60E-102.46E+053.93E-05#9104a-A67.68E-109.06E+046.96E-05#9104a-A73.54E-112.30E+058.13E-06#9104a-A82.37E-101.91E+054.51E-05#9104a-B11.37E-102.53E+053.45E-05#9104a-B21.14E-102.66E+053.03E-05#9104a-B33.51E-112.34E+058.23E-06#9104a-B41.53E-102.45E+053.74E-05#9104a-B5<1.0E-122.43E+05<1.0E-07#9104a-B61.14E-098.29E+049.41E-05#9104a-B71.63E-102.50E+054.07E-05#9104a-B82.80E-102.30E+056.44E-05#9104a-C11.39E-102.71E+053.76E-05#9104a-C21.72E-103.06E+055.28E-05#9104a-C31.76E-102.59E+054.55E-05#9104a-C42.03E-103.17E+056.42E-05#9104a-C51.92E-103.37E+056.46E-05#9104a-C69.28E-101.17E+051.09E-04#9104a-C71.52E-103.00E+054.56E-05#9104a-D19.48E-112.94E+052.79E-05#9104a-D21.49E-103.51E+055.22E-05#9104a-D32.29E-102.94E+056.73E-05#9104a-D42.06E-102.48E+055.09E-05#9104a-D51.30E-102.61E+053.39E-05#9104a-D61.07E-098.12E+048.70E-05#9104a-D71.98E-102.88E+055.69E-05#9104a-D81.77E-101.88E+053.33E-05#9104a-E12.8 9E-102.15E+056.21E-05#9104a-E22.12E-102.02E+054.27E-05#9104a-E32.80E-102.05E+055.74E -05#9104a-E41.97E-102.10E+054.15E-05#9104a-E52.73E-102.34E+056.36E-05#9104a-E61.19E- 096.55E+047.77E-05#9104a-E72.11E-102.18E+054.59E-05#9104a-E83.80E-101.49E+055.66E-05.

[0104]

[0105] As shown in Table 8, the manufactured antibodies were 10 -10 10 inland -9 It bound to human ASM protein with a binding affinity of M level.

Claims

1. An antibody or antigen-binding fragment thereof that specifically binds to ASM (acid sphingomyelinase) protein.

2. In the first paragraph, the ASM protein is an antibody or antigen-binding fragment thereof of mammalian origin.

3. In paragraph 1, the antibody or antigen-binding fragment thereof, A heavy chain variable region comprising a heavy chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 89, a heavy chain CDR2 comprising five or fewer amino acids substituted in a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 90, and a heavy chain CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 91; and An antibody or antigen-binding fragment thereof, comprising a light chain variable region comprising a light chain CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 92, a light chain CDR2 comprising a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 93 or wherein five or fewer amino acids are substituted in the polypeptide, and a light chain CDR3 comprising a polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 94 or wherein two or fewer amino acids are substituted in the polypeptide.

4. In the third paragraph, the heavy chain CDR2 in which five or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 90 is an antibody or antigen-binding fragment thereof in which five or fewer amino acids are substituted from the group consisting of the 2nd, 4th to 10th, 12th, 13th, and 17th amino acids from the N-terminus of the polypeptide.

5. In the third paragraph, the heavy chain CDR2 in which five or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 90 is an antibody or an antigen-binding fragment thereof, which is a polypeptide consisting of the amino acid sequence described in SEQ ID NOs: 95 to 111, respectively.

6. In the third paragraph, the light chain CDR2 in which five or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 93 is an antibody or antigen-binding fragment thereof in which four or fewer amino acids selected from the group consisting of amino acids 3 to 7 from the N-terminus of the polypeptide are substituted.

7. In the third paragraph, the light chain CDR2 in which five or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 93 is an antibody or an antigen-binding fragment thereof, which is a polypeptide consisting of the amino acid sequence described in SEQ ID NO: 112 to 135, respectively.

8. In the third paragraph, an antibody or antigen-binding fragment thereof, wherein the light chain CDR3 in which two or fewer amino acids are substituted in a polypeptide consisting of the amino acid sequence described in SEQ ID NO: 94, is an antibody or antigen-binding fragment thereof in which two or fewer amino acids are substituted from the group consisting of the 6th and 8th amino acids from the N-terminus of the polypeptide.

9. In the third paragraph, an antibody or antigen-binding fragment thereof, wherein the light chain CDR3 in which two or fewer amino acids are substituted in the polypeptide consisting of the amino acid sequence described in SEQ ID NO: 94 is a polypeptide consisting of the amino acid sequence described in SEQ ID NO:

136.

10. A nucleic acid encoding an antibody or an antigen-binding fragment thereof according to any one of claims 1 to 9.

11. An expression vector containing the nucleic acid of Article 10.

12. A host cell containing the nucleic acid of item 10 or the expression vector of item 11.

13. A method for producing an antibody or an antigen-binding fragment thereof that specifically binds to an ASM protein, comprising a step of culturing the host cell of item 12 to produce an antibody or an antigen-binding fragment thereof.