Characterization of Monoclonal Antibodies against sigma A Protein and Cross-Reactive Epitope Identification and Application for Detection of Duck and Chicken Reovirus Infections

文献类型： 外文期刊

第一作者： Chen, Xueming

作者： Chen, Xueming;Li, Tongtong;Chen, Xiaodan;Li, Chenxi;Lin, Weiwei;Liu, Hongyu;Song, Shuping;Bai, Xiaofei;Zhang, Yun

作者机构：

关键词： duck reovirus; avian reovirus; turkey reovirus; monoclonal antibody; sigma A protein epitopes; cross-reactive epitopes; diagnosis

期刊名称：PATHOGENS （ 影响因子：3.492； 五年影响因子：4.066 ）

ISSN：

年卷期： 2019 年 8 卷 3 期

页码：

收录情况： SCI

摘要： Although sigma A is an important major core protein of duck reovirus (DRV), the B-cell epitopes of this protein remain unknown to reseacrhers. Six monoclonal antibodies (MAbs) (1A7, 3F4, 5D2, 4E2, 3C7, and 2B7) were developed by using prokaryotic-expressed recombinant His-sigma A protein. Five of six MAbs (1A7, 3F4, 4E2, 3C7, and 2B7) reacted with His-sigma A protein in a conformation-independent manner, while 5D2 reacted with sigma A in a conformation-dependent manner. Immunofluorescence assays showed that the MAbs could specifically bind to DRV infected BHK-21 cells. The MAbs were delineated as three groups by a competitive binding assay. By using 12-mer peptide phage display and mutagenesis, MAb 4E2 was identified to recognize minimal epitope (56)EAPYPG(61) and MAb 1A7 recognize (WVV)-W-341/MAGLI/V-347, residues V-341/M and I-347/V are replaceable. Dot blotting and sequence analysis confirmed that EAPYPG and WVV/MAGLI/V are cross-reactive epitopes in both DRV and avian reovirus (ARV). An enzyme-linked immunosorbent assay (ELISA) based on two expressed EAPYPG and WVVAGLI as antigen demonstrated its diagnostic potential by specific reacting with serum samples from DRV- or ARV-infected birds. Based on these observations, an epitope-based ELISA could be potentially used for DRV or ARV surveillance. These findings provide insights into the organization of epitopes on sigma A protein that might be valuable for the development of epitope-based serological diagnostic tests for DRV and ARV infection.

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