Antibody capable of specifically binding to human β-klotho receptor and use thereof

A receptor-specific and antibody-based technology, applied in the field of genetic engineering, can solve the problem that the pharmaceutical properties of FGF21 cannot meet the requirements, etc.

Active Publication Date: 2021-02-09
郑州鸿运华宁生物医药有限公司
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  • Description
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  • Application Information

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Problems solved by technology

[0004] Although there are many examples of modifying natural FGF21 through genetic engineering to make it a new therapeutic drug (Adams et al., (2013) Plos One 8(6):e65763), the modified FGF21 is in some The pharmaceutical properties are far from meeting the requirements, such as pharmacokinetics, stability, solubility, etc.

Method used

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  • Antibody capable of specifically binding to human β-klotho receptor and use thereof
  • Antibody capable of specifically binding to human β-klotho receptor and use thereof
  • Antibody capable of specifically binding to human β-klotho receptor and use thereof

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Experimental program
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specific Embodiment

[0122] 1. Construction of stable antigen cell strains for immunization

[0123] Inoculate CHO-DHFR cells (Cell Bank of Chinese Academy of Sciences) into a 6-well plate, and after culturing for 24 hours, transfect the cells in the 6-well plate with a plasmid (commercially available) containing the V5-hKLB gene, and replace the culture medium before transfection. The transfection conditions of Lipofectamine 2000 are recommended for transfection. After 48 hours, change to the complete medium containing 10nM methotrexate (Methotrexate, MTX), and change the medium every 3 days. After about two weeks, stable growth clones appear, digest and disperse cell colonies, and wait until they grow to 50 When the % healing degree was obtained, the concentration of MTX was gradually increased for pressurized screening, and the highest concentration of MTX was 10uM. The constructed stable cell lines were detected by FACS, and the N'-terminal V5 tag antibody of KLB protein (FITC-anti-V5, Invitr...

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Abstract

The invention discloses an antibody capable of being specifically combined with human beta-Klotho receptor and application of the antibody. The antibody can be specifically combined with a co-receptor beta-Klotho of a fibroblast growth factor 21, is used for activating a human beta-Klotho / FGFR1c (Fibroblast Growth Factor Receptor 1c) complex receptor and can be used for treating or preventing type II diabetes mellitus and related diseases in mammals.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an antibody capable of specifically binding to human beta-Klotho receptors and its application. Background technique [0002] Fibroblast growth factor 21 (fibroblast growth factor 21, FGF21) is a new member of the fibroblast growth factor family (FGF19, FGF21 and FGF23), which is a secreted protein (Itoh et al., (2004) Trend Genet. 20:563-69). FGFs bind to growth factor receptors (FGFRs) present on the cell surface and transmit signals into the cell. There are four subtypes of FGFRs (FGFR1-4), which are transmembrane proteins and mainly consist of three parts: extracellular region, transmembrane region and intracellular region, including multiple alternative splicing bodies, such as FGFR1c, FGFR2c , FGFR3c and FGFR4. Like most growth factor receptors, FGFRs are tyrosine kinase receptors, which dimerize after binding to ligands, thereby activating tyrosine kinase ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N15/13A61K39/395A61P3/10
CPCC07K16/28C07K2317/56C07K2317/565C07K2317/92
Inventor 汪笑峰章华药晨江张成景书谦
Owner 郑州鸿运华宁生物医药有限公司
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