Production And Characterization Of Monoclonal Antibodies In Enterovirus71Diagnosis

Objective: As a highly neurotropic virus, EV71(enterovirus type71) has beenreported to be associated with several outbreaks throughout the Asia-Pacific region inrecent years and has become a major public health issue. Here, we work on generatingand characterizing EV71-specific monoclonal antibodies, and assay their potentialapplications in clinical diagnosis and antiviral study.Methods: BALB/c mice were immunized with EV71Tainan/4643/98strain. Apanel of hybridoma clones was selected for their secreting antibodies against EV71byIFA (immuno-fluorescence assay) of EV71-infected RD cells. We characterized theirisotypes, specificities and applications via flow cytometry, Western blot, antigen-captureELISA, and neutralization experiments. Their epitopes were further mapped bypeptide-ELISA. Applying these monoclonal antibodies, we confirmed the antiviraleffects of (-)-Epigallocatechin gallate (EGCG) against EV71in SH-SY5Y cell model.Results?(1) Total five hybridoma clones which secreted anti-EV71monoclonalantibodies were produced, named mAb21?mAb22?mAb24?mAb26and mAb27,respectively;(2) All five monoclonal antibodies can specifically react with EV71instead of the CVA16or ECHO6;(3) The applications in diagnosis of EV71infectioncould be carried out through IFA, flow cytometry, western blot and ELISA;(4) BothmAb22and mAb24harbor the neutralizing activities against EV71;(5) mAb21?mAb22?mAb24and mAb26recognize the conformation-dependent epitope within VP1,while the mAb27recognize the linear epitope (DVIESSIGDSVSRAL) located at theN-terminal of VP1;(6) Besides the potential values in EV71diagnostic, theseantibodies were also useful in antiviral studies as for screening of natural compoundsagainst EV71infection.Conclusions: We produced and characterized five anti-EV71monoclonal antibodies and further evaluated their applications in diagnosis of EV71and antiviralresearches.