Preparation and identification of antibodies against respiratory syncytial virus nucleoprotein / 中华微生物学和免疫学杂志

ABSTRACT

Objective To prepare and identify the monoclonal antibodies against respiratory syn-cytial virus nucleoprotein(RSV N protein). Methods A prokaryotic expression system was used to express recombinant RSV N protein in Escherichia coli (E.coli). BALB/c mice were immunized with the recombi-nant N protein after purification. Monoclonal antibodies (McAbs) against the N protein were sorted from these BALB/c mice and then were further characterized by Western blot, ELISA and immunofluorescence. Furthermore,this study used orthogonal experiment to identify McAbs pairs,which could be used for diagno-sis. Results This study succeeded in obtaining 24 hybridoma cells that stably secreted monoclonal antibod-ies against RSV N protein. These antibodies showed good reactivity in ELISA,of which eight had strong spe-cificity in Western blot and 13 could be used in immunofluorescence. This study obtained two McAbs pairs (12F2/11H8-HRP and 12F2/15A8-HRP) that could be used in RSV detection. Conclusion This study succeeded in screening and preparation of McAbs against RSV N protein and obtaining two potential McAbs pairs for rapid detection of RSV.