Preparation and Specificity Analysis of scFv Aganinst Salmonella enteritidis

Objective: To obtain the specific scFv against Salmonella enteritidis using genetic engineering technology.Methods: VH and VL genes were amplified by Reverse Transcription(RT) PCR from hybridoma cells secreting anti-Salmonella enteritidis monoclonal antibody,scFv gene(VL-Linker-VH) was obtained by Splice Overlap Extension(SOE) PCR with flexible polypeptide Linker connector(Gly4Ser)3.Subsequently the scFv-pGEX-4T-1 recombinant plasmid was constructed and transformed into E.coli BL21 for expression using IPTG as an inducer.The expressed recombinant protein was purified by GST chromatography and identified by SDS-PAGE and ELISA.Results: DNA sequencing demonstrated that scFv was successfully inserted into pGEX-4T-1.SDS-PAGE demonstrated that the molecular weight of the expressed scFv was about 60 kDa,and ELISA results confirmed that the obtained scFv can be recognized by not only Salmonella enteritidis but also S.enterica subsp enterzca,S.anatum and S.typhimurium.Conclusion: Specific scFv against S.enteritidis was obtained and can be used as candidate antibody molecules in detection of Salmonella.