DEVELOPMENT OF MONOCLONAL-ANTIBODIES AGAINST HANTAAN VIRUS NUCLEOCAPSID PROTEIN

Forty-five hybridoma cell lines producing monoclonal antibodies agains t Hantaan virus, the etiologic agent of hemorrhagic fever with renal s yndrome, were generated by fusion of P3-X63-Ag8.V653 myeloma cells wit h spleen cells of mice immunized with inactivated Hantaan virus vaccin e. Among these, 38 antibodies were identified as binding to the 48-kDa nucleocapsid protein by immunoblot assay or radioimmunoprecipitation. Twenty-six of them were of the immunoglobulin G1 (IgG1), nine were of the IgG2a, and three were of the IgA isotype. According to cross-reac tivities with other serotypes of the genus Hantavirus, the antibodies were classified into three groups: 6 antibodies specific to the Hantaa n serotype (group I), 20 antibodies crossreacting with Hantaan and Seo ul serotypes (SR-11, Tchoupitoulas, and R22) (group II), and 12 antibo dies cross-reacting with Hantaan, Seoul, and Prospect Hill serotypes ( group III). None of the antibodies crossreacted with the Puumala serot ype. With a panel of antibodies of different cross-reactivities, serot ypes of Hantavirus could be differentiated. Thirty-eight monoclonal an tibodies against Hantaan virus nucleocapsid protein which have differe nt cross-reactivities between serotypes were developed. These results confirmed the presence of multiple serotype-specific epitopes on the n ucleocapsid protein of Hantaan virus, which can be utilized in differe ntiation of serotypes.