Original Article
Construction of HIV-1 broadly neutralizing antibody gene eukaryotic expression vector and detection of its biological function
Wang Xiaofei, Jia Runqing, Zhou Yubai, Guan Yongjun, Zeng Yi
Published 2017-06-30
Cite as Chin J Exp Clin Virol, 2017,31(03): 198-201. DOI: 10.3760/cma.j.issn.1003-9279.2017.03.004
Abstract
ObjectiveTo construct a recombinant double gene co-expressing plasmid of HIV-1 broadly neutralizing antibody and to detect its expression in 293T cell line.
MethodsHIV-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.
ResultsThe 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.
ConclusionsThe 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.
Key words:
HIV; Neutralizing antibody; DNA vector
Contributor Information
Wang Xiaofei
Institute of Materia Medica, College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China
Jia Runqing
Institute of Materia Medica, College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China
Zhou Yubai
Institute of Materia Medica, College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China
Guan Yongjun
Institute of Materia Medica, College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China
Zeng Yi
Institute of Materia Medica, College of Life Science and Bioengineering, Beijing University of Technology, Beijing 100124, China; 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
