Background: To prepare monoclonal antibodies against West Nile virus (WNV) envelope protein domain.
Methods: BALB/c mice were immunized with recombinant antigen of West Nile virus envelope protein domain, and the spleen cells of the mice were used to prepare the monoclonal antibodies (McAb) by hybridoma technique.
Results: Three hybridoma cell strains secreting McAbs against WNV envelope protein domain, designated as 4F7, 6H3 and 8E4, respectively, were obtained and were identified by indirect enzyme linked immunosorbent assay (ELISA), they belonged to IgG1, IgG1 and Ig2a, respectively. Two epitopes of envelope protein domain were determined, among them, 4F7 and 6H3 were against the same epitope and 8E4 to another one.
Conclusions: The results of indirect ELISA, Western blot and indirect immunofluorescence experiment indicated that these three McAbs were specific for West Nile virus envelope protein domain and did not cross-react with Japanese encephalitis virus and other viruses, so they can be used for specific detection of West Nile virus.