Construction of HIV-1 broadly neutralizing antibody gene eukaryotic expression vector and detection of its biological function
10.3760/cma.j.issn.1003-9279.2017.03.004
- VernacularTitle: HIV-1中和抗体2G12真核表达载体的构建及活性检测
- Author:
Xiaofei WANG
1
;
Runqing JIA
1
;
Yubai ZHOU
1
;
Yongjun GUAN
1
;
Yi ZENG
1
,
2
Author Information
1. Institute of Materia Medica, College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China
2. State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China
- Publication Type:Journal Article
- Keywords:
HIV;
Neutralizing antibody;
DNA vector
- From:
Chinese Journal of Experimental and Clinical Virology
2017;31(3):198-201
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a recombinant double gene co-expressing plasmid of HIV-1 broadly neutralizing antibody and to detect its expression in 293T cell line.
Methods:HIV-1 neutralizing antibody 2G12 variable region of light chain (VL) and heavy chain (VH) was synthesized, and ligated with vectors containing human IgG constant regions of light and heavy chain to construct a complete 2G12 light and heavy Chain. The VL and VH of 2G12 and IRES were cloned into eukaryotic expression vector pVR by PCR amplification and then the recombinant plasmid was transfected into 293T cells. Expression of the antibody in cell supernatant was detected by ELISA. Binding and neutralizing activity of the cell supernatant were tested by ELISA and micro-neutralization assays.
Results:The recombinant double gene eukaryotic expression vector which can express human IgG was constructed successfully. The expression level of the supernatant was 6.43 μg/ml, and the antibody retained the binding and neutralizing activity.
Conclusions:The constructed vector can express the antibody with binding and neutralizing activity, this study provides a good platform for the expression of human HIV neutralizing antibody in the eukaryotic expression vector.
