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Anti human gas6 monoclonal antibody

A monoclonal antibody and antibody technology, applied in the direction of antibodies, antibody medical components, anti-animal/human immunoglobulin, etc., can solve problems such as unknown antibodies

Active Publication Date: 2017-09-08
KYOWA HAKKO KIRIN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Additionally, other Gas6 neutralizing antibodies are unknown

Method used

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  • Anti human gas6 monoclonal antibody
  • Anti human gas6 monoclonal antibody
  • Anti human gas6 monoclonal antibody

Examples

Experimental program
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Effect test

preparation example Construction

[0348] (1) Preparation of antigen

[0349] Human Gas6 used as an antigen can be obtained by purifying it from human Gas6 expression cells that have been introduced into E. coli, yeast, insect cells, or animal cells, and an expression vector containing a cDNA encoding the full length of human Gas6 or a partial length thereof. In addition, it can also be obtained by purifying human Gas6 from various human cell lines, human cells, human tissues, etc. that express human Gas6 in large quantities. Furthermore, a synthetic peptide containing a partial sequence of human Gas6 can also be prepared by chemical synthesis methods 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 also have known tags such as FLAG or His attached to the C-terminal or N-terminal.

[0350] The human Gas6 used in the present invention can use methods described in Molecular Cloning, A Laboratory Manual, Second Edition, Co...

Embodiment 1

[0492] [Example 1] Obtaining Gas6 knockout (hereinafter referred to as KO) mice

[0493] Sperm from Gas6 heterozygous KO mice was 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 was performed by CLEA Japan, Inc., the fertilized egg was transplanted into the recipient mouse to obtain the offspring. The obtained offspring were genotyped by a known method to confirm that the Gas6 gene has 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, recombinant human, cynomolgus, rat and mouse Gas6 proteins with FLAG tag attached to the C-terminus were prepared by the method described below. Hereinafter referred to as hGas6-F, cGas6-F, rGas6-F and mGas6-F.

[0496] (1) Construction of hGas6-F expression vector

[0497] From the plasmid (manufactured by Invitrogen) incorporating the gene sequence of human Gas6 (SEQ ID NO: 3, GenBank accession number: NM_000820), an animal cell expression vector incorporating the hGas6-F gene sequence was prepared in the following manner.

[0498] Using the above plasmid as a template, primers 1 and 2 (sequence numbers 1, 2) were used to amplify the DNA fragment containing the hGas6-F gene by polymerase chain reaction (PCR). In PCR, prepare a 20 μL reaction solution containing the template plasmid, 10 pmol each of the above two primers, and KOD FX (manufactured by Toyobo Co., Ltd.). After incubating...

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Abstract

The present invention pertains to a monoclonal antibody, or an antibody fragment thereof, that binds specifically to human growth arrest-specific 6 (hGas6) and binds to at least one amino acid residue among the 314th, 315th, and 316th amino acid residues of human Gas6. The present invention also pertains to: a nucleic acid containing a base sequence that codes for the antibody or the antibody fragment; a transformed cell containing a vector that contains the nucleic acid; a method for producing the antibody or the antibody fragment; reagents for detecting or measuring Gas6 that contain the antibody or the antibody fragment; therapeutic or diagnostic agents for Gas6-related diseases that contain the antibody or the antibody fragment as an active ingredient; and a use of the antibody or the antibody fragment for producing the therapeutic or diagnostic agents for Gas6-related diseases.

Description

Technical field [0001] [Associated Application] [0002] This specification includes the content described in the specification of U.S. Provisional Application US62 / 066,687 (filed on October 21, 2014) which is the basis of the 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 arrest specific protein 6 (hGas6), and is related to amino acid residues 314, 315 and 316 of human Gas6 At least one amino acid residue is bound; a nucleic acid containing a base sequence encoding the above-mentioned antibody or antibody fragment; a transformed cell containing the above-mentioned nucleic acid; a method for producing the above-mentioned antibody or antibody fragment; the detection or measurement of Gas6 using the above-mentioned antibody or antibody fragment Reagents; therapeutic drugs or diagnostic drugs for Gas6-related diseases containing the above-mentioned antibodies...

Claims

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Application Information

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IPC IPC(8): C12N15/09A61K39/395A61P13/12A61P35/00
CPCA61K39/39558C07K16/22A61K2039/505C07K2317/32C07K2317/33C07K2317/34C07K2317/73C07K2317/76A61P13/12A61P35/00A61P3/10C07K16/18G01N33/6893C07K2317/14G01N2333/4703A61K39/3955C07K16/28G01N33/53
Inventor 宇部优子山田刚士山崎雄司木村要播口晓子小川进也
Owner KYOWA HAKKO KIRIN CO LTD
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