[Preparation of monoclonal antibody against enterovirus 71 capsid protein VP1]

Objective To prepare murine monoclonal antibody (mAb) against enterovirus 71 (EV71) capsid protein VP1. Methods VP1 protein was expressed and purified. BALB/c mice were immunized with inactivated and purified EV71 virus, and mAb specific to EV71 VP1 was generated by hybridoma technique. Indirect ELISA was used to test antibody titer and antibody subclass identification. The expression of VP1 protein was detected by Western blot in EV71-infected RD cells. The expression and distribution of VP1 protein was detected by immunofluorescence cytochemistry in EV71-infected RD cells. Results Six antibody strains were obtained, among which three were IgG2a and three were IgG2b, all of which could be used for ELISA, Western blot and immunofluorescence cytochemical staining. 2D7 exhibited neutralization capacity with 50% inhibitory concentrations(IC₅₀) of 9.892 μg/mL. Conclusion Six strains of monoclonal antibodies with excellent reactivity were obtained, which laid a foundation for the further studies on the identification and diagnosis of EV71 as well as the functional of VP1 protein.