Screening and identification of linear B cell epitopes on the VP1 protein of enterovirus C96
Objective To identify linear B cell epitopes of VP1 protein of enterovirus C96(EV-C96).Methods The linear B cell epitopes and the property parameters of EV-C96 VP1 protein were predicted by the IEDB online analysis tools,combined with its secondary structure parameters such as β-turning angle,antigenicity,and hydrophilicity were used to screen out candidate epitopes with advantages.The reactivity of the candidate epitopes with the EV-C96 antiserum and the cross-reactivity with antiserums against 21 other common enterovirus caused hand foot and mouth disease were determined by indirect ELISA assay.The neutralization titer of epitope antibody was determined by microneutralization test.The conservativeness of epitopes was analyzed by sequence alignment,and the structural specificity of epitopes was analyzed by structural alignment.Results Seven epitopes of EV-C96 VP1 protein(P1～P7)were predicted by bioinformatics methods,among which P7(aa282～304)showed strong reactivity with EV-C96 antiserum and did not cross-react with other common enterovirus antisera.The neutralizing titer of P7 antibody was less than 1∶2.The amino acid sequence of P7 was highly conservative in EV-C96 isolates,and the structure of the P7 was different from other 21 common EVs.Conclusion P7 is a non-neutral EV-C96-specific B cell epitope that can be used as a candidate antigen epitope for the development of EV-C96 detection kits.
enterovirus C96VP1 proteinlinear B cell epitopeshand-foot-and-mouth disease
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