Antigenic analysis monoclonal antibodies against different epitopes of σB protein of Muscovy duck reovirus

摘要

σB is one of the major structural proteins of Muscovy duck reovirus (DRV), which is able to induce protective immune response in target birds. Four anti-DRV σB MAbs were identified belong to two distinct epitopes, designated A (1E5, 4E3, and 5D8) and B (2F7) (Liu et al., 2010). To understand antigenic determinants of the σB protein, a set of 20 (P1-P20), partially overlapping and consecutive peptides spanning σB were expressed and then screened by MAbs. With Western blot and enzyme-linked immunosorbent assay (ELISA), two minimal units of the linear epitopes, 19YIRAPACWD27 (epitope B) and 65TDGVCFPHHK74 (epitope A), were identified within N-terminal region of the σB protein. The epitope B was highly conserved among DRV and avian reovirus (ARV) strains through sequence alignment analysis. Immunofluorescence assays (IFA) and ELISA, confirmed that epitope B is a broad group-specific epitope among DRV and ARV. Epitope A could only react with chicken embyonated fibroblast cells (CEF) infected with DRV, but not ARV. However, both peptides have good immunogenicity and could induce antibodies against DRV in BALB/c mice. This report documents the first identification of σB epitopes in the precise locations. The two probes would be useful in the development of discriminating diagnostic kits for DRV and ARV infection.