Abstract
BackgroundRecently, 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.ResultsTwo 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.ConclusionThe 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.
Highlights
Serotype 4 fowl adenovirus (FAdV-4) has spread widely and caused huge economic loss to poultry industry
Two monoclonal antibody (mAb) specific to fiber-1 of Fowl adenovirus (FAdV)-4 were generated Balb/c mice were immunized with purified prokaryotic fusion protein GST-F1-S and LMH cells infected with FAdV-4 were used as a screening antigen for mAbs
The two mAbs showed great specificity to FAdV-C species, indicating mAb 3B5 and 6H9 could efficiently immunoprecipitate fiber-1 of FAdV-4 To further characterize the two mAbs generated here, LMH cells infected with FAdV-4 or transfected with pcDNA3.1-F1 were analyzed through Western blot and immunoprecipitation
Summary
Serotype 4 fowl adenovirus (FAdV-4) has spread widely and caused huge economic loss to poultry industry. The diseases caused by the infection of FAdV mainly include inclusion body hepatitis (IBH), hepatitis-hydropericardium syndrome (HPS), and gizzard erosion and ulceration (GEU) [2,3,4]. Since the commercial monoclonal antibody (mAb) against the fiber proteins of FAdV-4 is not available, the molecular mechanism for its infection and pathogenesis of FAdV-4 is largely unknown, and the efficient diagnostics for FAdV-4 is available. Two novel mAbs specific to the fiber-1 protein of FAdV-4 were generated. Both mAbs could efficiently immunoprecipitate the fiber-1 protein either in the infected cells or the transfected cells. A mAb based sandwich ELISA for specific detection of FAdV-4/10 was established
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