Cloning and characterization of the heavy chain variable region gene of monoclonal antibody 2F2 against Japanese encephalitis virus
10.3760/cma.j.issn.1008-1372.2014.05.001
- VernacularTitle:抗日本脑炎病毒单克隆抗体2F2重链可变区基因的克隆与鉴定
- Author:
Tiantian KOU
;
Yingfeng LEI
;
Tianbing DING
- Publication Type:Journal Article
- Keywords:
Encephalitis virus,Japanese;
Antibodies,monoclonal;
Cloning,molecular
- From:
Journal of Chinese Physician
2014;16(5):577-580
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain the heavy chain variable region (VH) gene of monoclonal antibody 2F2 against Japanese encephalitis virus (JEV).Methods Total RNA was isolated with Trizol from hybridoma 2F2 cells,and cDNA of VH was amplified with reverse transcription polymerase chain reaction (RT-PCR) and sequenced.The putative VH gene was expressed in E.coli,and the expressed products was detected with enzyme linked immunosorbent assay (ELISA) to determine the activity to bind JEV.Results The VH gene was 354 bp in length which encodes 118 amino acids.Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the VH gene was successfully expressed and purified from inclusion bodies.ELISA result also demonstrated that VH gene expressed products bind purified JEV.Conclusions The VH gene of monoclonal antibody 2F2 against JEV had been cloned.
