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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Publications at this Location » Publication #381584

Research Project: Non-antibiotic Strategies to Control Enteric Diseases of Poultry

Location: Animal Biosciences & Biotechnology Laboratory

Title: Development of antigen sandwich ELISA to detect interferon-alpha (IFN-a) using monoclonal antibodies in chicken

item LEE, YOUNGSUB - US Department Of Agriculture (USDA)
item KIM, WOOHYUN - Gyeongsang National University
item Lillehoj, Hyun

Submitted to: American Association of Immunologists Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 4/1/2021
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Interferon alpha (IFN-a) belongs to the type I interferon family which mediates an early innate immune response to viral infections. In the present study, we developed sandwich ELISA using specific mouse monoclonal antibodies (mAbs) to measure IFN-a production in chickens. Recombinant chicken IFN-a (chIFN-a) expressed in yeast was used to immunize the mice. Five mAbs which specifically recognize chicken IFN-a antigen were selected and characterized. For sandwich ELISA development, mAbs were labeled with biotin, followed by a pairing test to identify the best capture and detection antibodies. Two sets of mouse anti-chIFN-a mAb pairs were identified as optimum pairs of antibodies for antigen capture assay for detection of chIFN-a. The sandwich ELISA effectively detected a native chicken IFN-a in chicken macrophage cells stimulated by polyinosinic:polycytidylic acid (poly I:C), and its minimum detectable level was about 25 pg/mL. The anti-viral activity of chIFN-a against vesicular stomatitis virus was evaluated in avian embryonic fibroblast and the mouse anti-chIFN-a mAbs which neutralize functional activity of chicken IFN-a were identified. The newly developed antigen capture sandwich ELISA for the detection of chIFN-a will be a useful tool to monitor IFN-a production in chickens during viral infections.