Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (10): 179-185.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1525

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Preparation and Identification of a Novel FGF20 Monoclonal Antibody

TANG Lu1,2(), DONG Li-ping1, YIN Mo-li1,2, LIU Lei1,2, DONG Yuan1, WANG Hui-yan1,2()   

  1. 1. Jilin Medical University,Jilin 132013
    2. Jilin Collaborative Innovation Center for Antibody Engineering,Jilin 132013
  • Received:2020-12-17 Online:2021-10-26 Published:2021-11-12
  • Contact: WANG Hui-yan E-mail:770453679@qq.com;zswhy518@163.com

Abstract:

Prokaryotic expression vector for expression recombinant fibroblast growth factor 20(FGF20)was constructed,and monoclonal antibody against FGF20 was prepared,which may lay foundation for developing novel medicines. The prokaryotic expression vector pET24a-SUMO-hFGF20 was constructed and transformed into Escherichia coli BL21(DE3)for induction and expression. The fusion protein was purified by Ni NTA affinity chromatography column,digested with SUMO enzyme,and purified again by Ni NTA affinity chromatography column. The purity of the protein was detected by SDS-PAGE. Balb/c mice were immunized with the recombinant FGF20 protein,and the monoclonal antibody was prepared by conventional way,and isolated and purified by protein G column. The purity of the antibody was detected by SDS-PAGE. The titer and subtype of the antibody were detected by ELISA. The affinity of the antibody was determined by Western blot and SPR. The recombinant plasmid pET24a-SUMO-hFGF20 was successfully constructed. The relative molecular weight of the isolated and purified FGF20 protein was 23 kD. A highly positive hybridoma cell line detected by ELISA was obtained. After purification,the titer of the monoclonal antibody was 1:25 600,and the subtype is IgG1. It bound to commercial antigen,and the affinity was 1.07×10 -7 M. High-affinity monoclonal antibody against the FGF20 protein was successfully prepared by construction,expression and purification of recombinant protein.

Key words: fibroblast growth factor 20(FGF20), prokaryotic expression, protein purification, monoclonal antibody