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Anti-human gas6 monoclonal antibody
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A monoclonal antibody, antibody technology, applied in the direction of antibodies, antibody medical components, anti-animal/human immunoglobulins, etc., can solve the problem of unknown antibodies and other issues
Active Publication Date: 2021-07-20
KYOWA HAKKO KIRIN CO LTD
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Additionally, other Gas6 neutralizing antibodies are unknown
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[0349] Human Gas6 as an antigen can be obtained by purifying human Gas6-expressing cells into which an expression vector containing cDNA encoding the full-length or partial length of human Gas6 has been introduced into Escherichia coli, yeast, insect cells, animal cells, or the like. In addition, human Gas6 can also be obtained by purifying human Gas6 from various human cell lines, human cells, human tissues, etc. expressing a large amount of human Gas6. Furthermore, a synthetic peptide containing a partial sequence of human Gas6 can be prepared by a chemical synthesis method such as the Fmoc method or the tBoc method, and used as an antigen. Human Gas6 or a synthetic peptide having a partial sequence of human Gas6 may have a known tag such as FLAG or His added to the C-terminus or N-terminus.
[0350] The human Gas6 used in the present invention can use methods described in Molecular Cloning, A Laboratory Manual, Second Edition, Cold Spring H...
Embodiment 1
[0492] [Example 1] Acquisition of Gas6 knockout (hereinafter referred to as KO) mice
[0493] Sperm from Gas6 heterozygous KO mice were purchased from Taconic. The background of the purchased Gas6 heterozygous KO mice was 129 / SvEv-C57BL / 6. After in vitro fertilization of the sperm of the Gas6 heterozygous KO mouse and the egg of the C57BL / 6NJcl mouse by CLEA Japan, Inc., the fertilized egg was transplanted into the recipient mouse to obtain offspring. The obtained pups were genotyped by a known method, and it was confirmed that the Gas6 gene had been knocked out, and Gas6 homozygous KO mice were obtained.
Embodiment 2
[0494] [Example 2] Production of various Gas6 recombinants
[0495] For immunization and screening, human, cynomolgus monkey, rat, and mouse Gas6 recombinant proteins with FLAG tag added to the C-terminus were produced by the methods described below. Hereinafter, they are referred to as hGas6-F, cGas6-F, rGas6-F, and mGas6-F, respectively.
[0497] From a plasmid (manufactured by Invitrogen) incorporating the human Gas6 gene sequence (SEQ ID NO: 3, GenBank accession number: NM_000820), an animal cellexpression vector incorporating the hGas6-F gene sequence was prepared as follows.
[0498] Using the above plasmid as a template, a DNA fragment containing the hGas6-F gene was amplified by polymerasechain reaction (PCR) using primers 1 and 2 (SEQ ID NO: 1, 2). In PCR, 20 μL of a reaction solution containing a template plasmid, 10 pmol each of the above two primers, and KOD FX (manufactured by Toyobo Co., Ltd.) wa...
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Abstract
The present invention relates to: a monoclonalantibody or the antibody fragment, which is an antibody that specifically binds to human growth arrestspecific protein 6 (hGas6), and the 314th, 315th and 316th amino acid residues of human Gas6 Combination of at least one amino acid residue; a nucleic acid containing a base sequence encoding the above-mentioned antibody or antibody fragment; a transformed cell comprising a vector containing the above-mentioned nucleic acid; a method for producing the above-mentioned antibody or antibody fragment; Gas6 containing the above-mentioned antibody or antibody fragment Reagents for detection or measurement; therapeutic or diagnostic drugs for Gas6-related diseases containing the above-mentioned antibodies or antibody fragments as active ingredients; or, applications of the above-mentioned antibodies or antibody fragments in the manufacture of therapeutic or diagnostic drugs for Gas6-related diseases.
Description
technical field [0001] [Association Application] [0002] This specification includes the content described in the specification of US provisional application US62 / 066,687 (filed on October 21, 2014) which is the basis of priority of this application. The present invention relates to: a monoclonal antibody or the antibody fragment, which is an antibody that specifically binds to human growth arrestspecific protein 6 (hGas6), and the 314th, 315th and 316th amino acid residues of human Gas6 Combination of at least one amino acid residue; nucleic acid containing a nucleotide sequence encoding the above-mentioned antibody or antibody fragment; transformed cell containing the above-mentioned nucleic acid; method for producing the above-mentioned antibody or antibody fragment; detection or measurement of Gas6 using the above-mentioned antibody or antibody fragment Reagents; therapeutic or diagnostic drugs for Gas6-related diseases containing the above-mentioned antibodies or antib...
Claims
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