Abstract:

To prepare monoclonal antibodies (Mabs) against capsid protein of swine hepatitis E virus Ⅳ (HEV), the gene of swine HEV encoding Cterminal 267 (408-675) amino acids of capsid protein was cloned into pET-28a (+) vector to construct recombinant plasmid pET-28a-ORF2-C. The protein was expressed in E. coli Rosetta (BL21), and identified by SDS-PAGE and Western blot. Spleens of BALB/c mice immunized with the purified protein were used to produce monoclonal antibodies (Mabs). The Mabs were selected and characterized with the indirect ELISA and competitive ELISA. Protein was expressed correctly. Three Mabs, designated Mab-2C7, 2G9 and 1E4, were selected and recognized three independent epitopes. Mab-2G9 is IgG2b and Mab-2C7 and Mab-1E4 are IgG1. The positive HEV swine sera could be blocked by Mab-1E4 and Mab-2G9 indicating the epitopes recognized by Mab-1E4 and Mab-2G9 are predominant epitopes in capsid protein of swine HEV genotype Ⅳwhile Mab-2C7 can not. The results suggest that the Mab-1E4 and Mab-2G9 are potential diagnostic reagents for detection of swine HEV infection genotype Ⅳ.