Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2017-0228

Abstract

Abstract: The aims of this study are to establish a method for efficient soluble expression of human squamous cell carcinoma antigen（SCCAg）based on Escherichia coli expression system and obtain the recombinant SCCAg antigen in fine activity,then apply it in the detection method establishment of antigen. The study on the method of soluble expression and purification of recombinant SCCAg antigen was conducted based on pGEX-6P-1 vector and E. coli ER2566 strain. The activity of the purified antigen was evaluated by Abbott Kit and the specific monoclonal antibody was screened by indirect ELISA. It was proved that PGEX-6P-1 vector and E. coli strain ER2566 could be used to establish efficient soluble expression and purification method for recombinant SCCAg antigen. Moreover,the recombinant SCCAg antigen was proved to be in high purity and activity. Thus,the SCCAg detection method of chemical luminous tube was established with the specific monoclonal antibodies. In conclusion,an effective method for the expression and purification of SCCAg,which is based on the E. coli expression system,is established.

Key words: human squamous cell carcinoma antigen, Escherichia coli expression system, soluble expression, monoclonal antibody

CHEN Chun-ye,LIU Jian,ZHU Rui,LI Shu-xuan,YE Jiang-hui,WANG Wei,PAN De-quan,XU Fei-hai,CHENG Tong,XIA Ning-shao. Preparation and Application of Soluble Human Squamous Cell Carcinoma Antigen Expressed by Escherichia coli[J]. Biotechnology Bulletin, 2017, 33(9): 252-258.

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