Aim: To prepare the monoclonal antibodies (mAbs) against Norovirus capsid protein for the development of a rapid assay of Norovirus and to investigate the pathogenesis of this virus.
Methods: Sp2/0-Ag-14 myeloma cells were fused with spleen cells of BALB/c mice immunized with the recombinant protein of Norovirus NVgz01 (DQ369797), which was overexpressed in E.coli. The monoclonal antibodies against Norovirus capsid protein were screened by selective culture medium. The obtained Ig subtypes, titer and specificity of the mAbs were examined by ELISA and Western blot respectively.
Results: After cell fusion and subcloning, four hybridoma cell lines which secreted monoclonal antibodies specifically against Norovirus capsid protein were obtained and named as N2C3, N7C2, N4B1 and N8A9. Indirect ELISA and Western blot assay showed that the four mAbs specifically recognized the Norovirus capsid protein expressed in E.coli. The native Norovirus capsid protein in the stool samples were also recognized by them. The Sandwich ELISA, a rapid detection assay of the expressed and native Norovirus capsid protein, demonstrated that N2C3 and N7C2 were matched successfully.
Conclusion: The mAbs against GII Norovirus capsid protein have been developed, which can be used for the development of detection assay and the basic research of Norovirus.