Identification of novel B cell epitopes in Fiber-2 protein of duck adenovirus 3 and their application

AMB Express

Original Article

Wenyuan Zhang¹  Weikang Wang¹  Zhimin Wan¹  Jianqiang Ye¹  Quan Xie¹  Hongxia Shao¹  Aijian Qin¹  Yun Lin¹  Jing Xie¹  Tuofan Li¹ 

[1] Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine, Ministry of Education, College of Veterinary Medicine, Yangzhou University, 225009, Yangzhou, Jiangsu, China;Jiangsu Co-innovation Centre for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, China;Joint International Research Laboratory of Agriculture and Agri-Product Safety, the Ministry of Education of China, Yangzhou University, 225009, Yangzhou, Jiangsu, China;

关键词: Duck adenovirus 3;    Fiber-2 protein;    Monoclonal antibodies;    Epitope mapping;    Sandwich ELISA;   

DOI  :  10.1186/s13568-023-01552-9

 received in 2022-06-14, accepted in 2023-04-24,  发布年份 2023

来源: Springer

【 摘 要 】

Duck adenovirus 3 (DAdV-3), a newly emerged duck adenovirus, has resulted in significant economic losses to the duck industry across China since 2014. However, little is known about the B cell epitopes in major antigen of DAdV-3 and the serological approach for detection of DAdV-3 is not available. In this study, four monoclonal antibodies (mAbs) specific to Fiber-2 protein of DAdV-3 were first generated and designated as 2G10, 3D9, 5E6, and 6B12. Indirect immunofluorescence assay (IFA) showed that all of the mAbs reacted with the Fiber-2. Moreover, mAbs 2G10, 5E6, and 6B12 demonstrated good activity with Fiber-2 in Western blot. Notably, the Fiber-2 could be immunoprecipitated efficiently by mAb 3D9. Epitope mapping revealed that mAbs 2G10, 3D9, 5E6, and 6B12 recognized 397-429aa, 463-481aa, 67-99aa, and 1-66aa of Fiber-2, respectively. Besides, a novel sandwich ELISA for efficient detection of DAdV-3 was developed based on mAb 3D9 and horseradish peroxidase (HRP) conjugated mAb 3D9. The sandwich ELISA only reacted with DAdV-3 but not with other duck-associated viruses. The limit of detection of the ELISA was 6.25 × 103 TCID50/mL. Overall, the mAbs generated laid the foundation for elucidating the critical role of Fiber-2 in mediating infection and pathogenesis, and the sandwich ELISA approach established here provided efficient and rapid serological diagnostic tool for DAdV-3.

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

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