title: Generation and Characterization of a scFv Antibody Against T3SS Needle of Vibrio parahaemolyticus.
abstract: Vibrio parahaemolyticus, a halophilic gram-negative bacterium, is a food-borne pathogen that largely inhabits marine and estuarine environments, and poses a serious threat to human and animal health all over the world. The hollow "needle" channel, a specific assemble of T3SS which exists in most of gram-negative bacteria, plays a key role in the transition of virulence effectors to host cells. In this study, needle protein VP1694 was successfully expressed and purified, and the fusion protein Trx-VP1694 was used to immunize Balb/c mice. Subsequently, a phage single-chain fragment variable antibody (scFv) library was constructed, and a specific scFv against VP1694 named scFv-FA7 was screened by phage display panning. To further identify the characters of scFv, the soluble expression vector pACYC-scFv-skp was constructed and the soluble scFv was purified by Ni(2+) affinity chromatography. ELISA analysis showed that the scFv-FA7 was specific to VP1694 antigen, and its affinity constant was 1.07 × 10(8 )L/mol. These results offer a molecular basis to prevent and cure diseases by scFv, and also provide a new strategy for further research on virulence mechanism of T3SS in V. parahaemolyticus by scFv.
authors.affiliation: The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 China.
authors.forename: Rongzhi
authors.initials: R
authors.lastname: Wang
authors.affiliation: The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 China.
authors.forename: Sui
authors.initials: S
authors.lastname: Fang
authors.affiliation: The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 China.
authors.forename: Shuangshuang
authors.initials: S
authors.lastname: Xiang
authors.affiliation: The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 China.
authors.forename: Sumei
authors.initials: S
authors.lastname: Ling
authors.affiliation: The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yuan
authors.affiliation: The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002 China.
authors.forename: Shihua
authors.initials: S
authors.lastname: Wang
keywords: Biopanning; Needle; T3SS; Vibrio parahaemolyticus; scFv