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

State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150069, China
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Author to whom correspondence should be addressed.
Authors contribute equally.
Pathogens 2019, 8(3), 140; https://doi.org/10.3390/pathogens8030140
Received: 6 August 2019 / Revised: 29 August 2019 / Accepted: 30 August 2019 / Published: 7 September 2019
(This article belongs to the Section Animal Pathogens)
Although σA 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-σA protein. Five of six MAbs (1A7, 3F4, 4E2, 3C7, and 2B7) reacted with His-σA protein in a conformation-independent manner, while 5D2 reacted with σA 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 56EAPYPG61 and MAb 1A7 recognize 341WVV/MAGLI/V347, residues 341V/M and 347I/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 σA protein that might be valuable for the development of epitope-based serological diagnostic tests for DRV and ARV infection. View Full-Text
Keywords: duck reovirus; avian reovirus; turkey reovirus; monoclonal antibody; σA protein epitopes; cross-reactive epitopes; diagnosis duck reovirus; avian reovirus; turkey reovirus; monoclonal antibody; σA protein epitopes; cross-reactive epitopes; diagnosis
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MDPI and ACS Style

Chen, X.; Li, T.; Chen, X.; Li, C.; Lin, W.; Liu, H.; Song, S.; Bai, X.; Zhang, Y. Characterization of Monoclonal Antibodies against σA Protein and Cross-Reactive Epitope Identification and Application for Detection of Duck and Chicken Reovirus Infections. Pathogens 2019, 8, 140.

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