禽腺病毒血清4型Fiber2蛋白单克隆抗体制备及其抗原表位鉴定

doi:10.11843/j.issn.0366-6964.2021.07.022

Abstract

Abstract: To prepare monoclonal antibodies specific to FAdV-4 Fiber2 protein, this study used NusA-Fiber2, a soluble recombinant protein expressed in prokaryotes, as an immunogen to immunize BALB/c female mice. Three hybridomas (2G5, 2G8 and 4C2) secreting monoclonal antibodies (MAb) against FAdV-4 Fiber2 protein were selected. Ascites were prepared from cell line 2G5 and purified. The specificity of the monoclonal antibody was determined by indirect immunofluorescence assay and Western blot. The prepared monoclonal antibody was used to coat the enzyme plate. Through a series of optimization, the detection method of FAdV-4 Fiber2 double antibody sandwich ELISA was established. The epitopes recognized by monoclonal antibodies were identified by progressively truncating Fiber2 protein. Results were as follows:Three hybridomas 2G5, 2G8 and 4C2 were successfully obtained. IFA and Western blot results showed that MAb 2G5 could react specifically with purified Fiber2 protein and FAdV-4 virus. The results showed that the sandwich ELISA was specific, sensitive and reproducible. Our results revealed that the antigenic epitope required for reactivity with the 2G5 was N-terminal aa1-33. In this study, monoclonal antibodies with good reactivity of Western blot and IFA were successfully prepared. This provides experimental data and theoretical basis for the development of rapid serological diagnosis and commercial vaccine against FAdV-4.

Key words: fowl adenovirus serotype 4 (FAdV-4), Fiber2 protein, monoclonal antibodies, sandwich ELISA, epitope identification

CLC Number:

S852.659.1

GUO Ruizhen, SU Bingqian, WANG Qi, WANG Yi, YU Pengwei, MENG Jiejie, QI Yanli, YANG Guoyu, CHU Beibei. Preparation of the Monoclonal Antibody against the Serotype 4 Fowl Adenovirus Fiber2 Protein and Identification of the Antigenic Epitope[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(7): 2000-2012.

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Preparation of the Monoclonal Antibody against the Serotype 4 Fowl Adenovirus Fiber2 Protein and Identification of the Antigenic Epitope

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