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Two novel monoclonal antibodies against fiber-1 protein of FAdV-4 and their application in detection of FAdV-4/10

Shao, Hongxia, Lu, Yanan, Wang, Weikang, Li, Tuofan, Zhang, Jianjun, Wan, Zhimin, Liang, Guangchen, Gao, Wei, Qin, Aijian, Ye, Jianqiang
BMC veterinary research 2019 v.15 no.1 pp. 232
Adenoviridae, detection limit, diagnostic techniques, enzyme-linked immunosorbent assay, epitopes, financial economics, fluorescent antibody technique, monoclonal antibodies, pathogenesis, poultry industry, serotypes, veterinary medicine
BACKGROUND: Recently, serotype 4 fowl adenovirus (FAdV-4) has spread widely and caused huge economic loss to poultry industry. However, little is known about the molecular pathogenesis of FAdV-4. Fiber protein is thought to be vital for its infection and pathogenesis. RESULTS: Two novel monoclonal antibodies (mAbs) targeting the fiber-1 protein of FAdV-4 were generated, designated as mAb 3B5 and 6H9 respectively. Indirect immunofluorescence assay (IFA) showed that both mAbs only reacted with the FAdV-4 and FAdV-10, not with other serotypes including FAdV-1, FAdV-5, FAdV-6, FAdV-7, FAdV-8 and FAdV-9 tested. Although both mAbs did not recognize the linear epitopes, they could efficiently immunoprecipitate the fiber-1 protein in LMH cells either infected with FAdV-4 or transfected with pcDNA3.1-Fiber-1. Moreover, mAb 3B5 as a capture antibody and HRP-conjugated mAb 6H9 as a detection antibody, a novel sandwich ELISA for efficient detection of FAdV-4 was generated. The limit of detection of the ELISA could reach to 1000 TCID₅₀/ml of FAdV-4 and the ELISA could be efficiently applied to detect FAdV-4 in the clinical samples. CONCLUSION: The two mAbs specific targeting fiber-1 generated here would pave the way for further studying on the role of fiber-1 in the infection and pathogenesis of FAdV-4, and the established mAb based sandwich ELISA would provide an efficient diagnostics tool for detection of FAdV-4/10.