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Human source Fab antibody for anti recombined alkalescent fibroblast growth factor and application

A technology of fibroblasts and growth factors, applied in anti-growth factor immunoglobulins, antibodies, anti-animal/human immunoglobulins, etc., can solve the problem of not seeing anti-bFGF human antibodies

Inactive Publication Date: 2007-12-26
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The anti-tumor angiogenesis and anti-tumor effects of bFGF monoclonal antibody have been confirmed and affirmed by scholars at home and abroad, but so far, there are no other domestic and foreign literature and patent reports on anti-bFGF human antibody research

Method used

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  • Human source Fab antibody for anti recombined alkalescent fibroblast growth factor and application
  • Human source Fab antibody for anti recombined alkalescent fibroblast growth factor and application
  • Human source Fab antibody for anti recombined alkalescent fibroblast growth factor and application

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Experimental program
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Effect test

Embodiment 1

[0083] Construction method of anti-recombinant basic fibroblast growth factor (bFGF) or fibroblast growth factor 2 (FGF-2) human Fab antibody:

[0084] 1. Cloning of Fd chain and κ chain genes and construction of κ chain library

[0085] Lymphocytes were isolated from peripheral blood of patients with high titer of bFGF or (FGF-2) autoantibody. Total cellular RNA was extracted with Trizol (TAKARA company, RNA extraction reagent) reagent, and cDNA was synthesized by reverse transcription with Olig-dt primer and ReverTra Ace-a (see TOYOBO company kit product manual) kit. Use a set of specific IgG Fab Fd and κ chain primers (see Table 1) (Fd-6 chain 3' primer is VH2 and 5' primer is VH6; κ-6 chain 3' primer is Vκ1, κ-6 chain 5 'The primer is Vκ3) to perform PCR amplification on the Fd and κ chains of the human Fab (see Table 2 for the reaction system). ] × 29, 72°C for 10 min.

[0086] Table 1 Primers for PCR amplification of human antibody Fab fragment genes

[0087] ...

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Abstract

This invention discloses anti-recombinant alkaline fibroblast growth factor human-derived Fab antibody and its application. The antibody segment genome comprises: heavy chain Fd chain and light chain kappa chain. This invention utilizes phage surface display technique to successfully construct kappa chain and Fd chain genes of human-derived Fab antibody that can neutralize recombinant human bFGF or FGF-2. Functional neutralizing antibody that can specifically bind bFGF or FGF-2 is obtained in prokaryotic cells, yeast cells, insect cells and eukaryotic cells, which can be used to prepare antibody drugs that can diagnose and treat tumors, and inhibit kidney, liver and lung fibrosis. The bFGF or FGF-2 human-derived Fab antibody has high affinity and specificity, and can be directly used to develop antibody drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an anti-recombinant basic fibroblast growth factor (or called fibroblast growth factor 2, FGF-2) human Fab antibody and application thereof. Background technique [0002] Basic fibroblast growth factor (bFGF), also known as fibroblast growth factor 2 (fibroblast growth factor 2, FGF-2), is a member of the fibroblast growth factor (fibroblast growth factor, FGF) family currently known The earliest and most studied factor among the 23 members of . bFGF has a variety of biological effects, including growth-promoting, division, and differentiation functions; it has a strong pro-proliferation effect on fibroblasts. [0003] bFGF is one of the most important cytokines stimulating angiogenesis after VEGF. It is closely related to the occurrence and development of tumors. bFGF is involved in the growth of various tumors (such as glioma, rhabdomyoma, leukemia, kidney cancer, lun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/22A61K39/395A61P35/00A61P43/00
Inventor 邓宁向军俭王宏唐勇汤伟佳关文答陶俊
Owner JINAN UNIVERSITY
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