鸡传染性贫血病毒VP2蛋白单克隆抗体的研制及其识别抗原表位的鉴定

doi:10.11843/j.issn.0366-6964.2022.02.026

Abstract

Abstract: The aim of this study was to prepare monoclonal antibody(mAb) against VP2 protein of chicken infectious anemia virus (CIAV) for diagnosis of CIAV and further research in characteristics of CIAV. The VP2 gene of CIAV was amplified and cloned into the pET-32a plasmid. The VP2 fusion protein was expressed in Escherichia coli induced by IPTG. Hybridoma cells were fused between SP2/0 cells and spleen cells of BALB/c mice immunized with the expressed protein and screened by immunofluenscence assay. The epitope recognized by mAb was further identified by the truncated VP2 proteins in Western blot. One hybridoma cell line named CIAV-VP2-4A12 that can stably secrete mAb against CIAV VP2 protein was successfully obtained. The isotype of the mAb was IgG1 and the light chain was kappa chain. Western blot assay showed that mAb could react with the VP2 protein expressed in Escherichia coli, Sf9 cells or DF1 cells transfected with pCAGSS-VP2-Flag. The epitope recognized by the mAb CIAV-VP2-4A12 was identified as ¹⁵⁵KTVRW¹⁵⁹of VP2, which sequence is highly conserved among different CIAV strains. In this study, a monoclonal antibody against CIAV VP2 was successfully developed and the antigenic epitope recognized by the mAb was identified, which will be useful tool for research on the function of the VP2 protein and development of diagnostic methods for CIAV.

Key words: chicken infectious anemia virus, VP2 protein, monoclonal antibody, epitope

CLC Number:

WANG Mingrui, WU Junhua, WANG Fei, SHAO Hongxia, QIAN Kun, YE Jianqiang, QIN Aijian. Development and Epitope Identification of a Monoclonal Antibody against VP2 Protein of Chicken Infectious Anemia Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 597-606.

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Development and Epitope Identification of a Monoclonal Antibody against VP2 Protein of Chicken Infectious Anemia Virus

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