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

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• 作者：Yongfeng Li ; Xiuchen Yin ; Xiaodan Chen ; Xiaojun Li ; Jinzhe Li ; Chunguo Liu ; Ming Liu ; ^{liuming04@126.com} ; Yun Zhang ; ^{yunzhang03@yahoo.com}
• 关键词：Muscovy duck reovirus (DRV) ; ¦ÒB protein ; Pepscan ; Epitope
• 刊名：Virus Research
• 出版年：2012
• 出版时间：February 2012
• 年：2012
• 卷：163
• 期：2
• 页码：546-551
• 全文大小：689 K

文摘

¦ÒB 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 ¦ÒB MAbs were identified belong to two distinct epitopes, designated A (1E5, 4E3, and 5D8) and B (2F7) (). To understand antigenic determinants of the ¦ÒB protein, a set of 20 (P1-P20), partially overlapping and consecutive peptides spanning ¦ÒB 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 ¦ÒB 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 ¦ÒB epitopes in the precise locations. The two probes would be useful in the development of discriminating diagnostic kits for DRV and ARV infection.