Cloning and characterization of the heavy chain variable region gene of monoclonal antibody 2F2 against Japanese encephalitis virus / 中国医师杂志

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.

Screening of mimicry peptide of Japanese encephalitis virus E protein from phage 15-mer peptide library / 细胞与分子免疫学杂志

Aim To study the mimicry peptide of JEV E protein by screening a phage 15-mer peptide library with anti-JEV E protein mAb 2H4. Methods After three rounds biopanning, the enriched positive phage clones were identified by ELISA. 10 positive phage clones were sequenced and compared homologously with JEV E protein. Results The short peptide displayed on screened positive phage could bind specifically to mAb 2H4, and the binding could be inhibited by natural JEV Ag. Amino acid sequences of the 10 positive phage clones were consensus, that is, RQDPQWPYANSTIAR. By homology analysis, two higher homologous sequences STXAR and WXXAXST were found in different regions of JEV E protein. The peptide displayed on positive phage could react specifically with the mouse antiserum against natural JEV Ag . Conclusion This peptide displayed on positive phage may mimic partial antigenicity of JEV E protein.