Development of murine monoclonal antibodies with potent neutralization effects on enterovirus 71

Enterovirus 71 (EV71) is a member of the Picornaviridae family and one of the main causative agents of hand, foot, and mouth disease (HFMD). Currently, EV71 infection is prevalent in the Asia-Pacific regions where it affects millions of children under the age of five, causing significant morbidity and mortality. No specific vaccine or antiviral drugs are available for EV71. The development of murine monoclonal antibodies (mAbs) with potent neutralization effects on EV71 is described. Mab-secreting hybridomas were generated from mice immunized with EV71 recombinant virus-like particles. Three IgG1 mAbs, D5, H7, and C4, capable of binding to and neutralizing EV71, were identified. In ELISA and Western blot assays, these mAbs reacted with recombinant VP1 protein, but not with VP0. They also detected cells infected with EV71 by immunofluorescent staining. In addition, these three mAbs had potent EV71 neutralization capacity, with 95% inhibitory concentrations of 0.3125, 0.3125, and 1.25 μg/ml for D5, H7, and C4, respectively. The presented data demonstrate that the anti-EV71 mAbs are not only promising candidates for development into humanized mAb for treatment but also useful reagents for development of diagnostic tests.