非洲猪瘟病毒p62蛋白单克隆抗体的制备及初步应用

doi:10.11843/j.issn.0366-6964.2020.05.018

Abstract

Abstract: The objective of this study was to prepare monoclonal antibodies against p62 protein of African swine fever virus (ASFV), and apply it to immunohistochemistry (IHC) detection of ASFV antigen in infected tissue samples. BALB/c mice were immunized with purified recombinant p62 protein expressed by baculovirus system and hybridoma cells were obtained by fusion of spleen cells and myeloma cells. An indirect ELISA based on the purified p62 protein was developed. Eighteen hybridoma cell lines stably secreting MAbs against the p62 protein were harvested after screening and subcloning. The results of indirect immunofluorescence assay (IFA) showed that all of the monoclonal antibodies could react with ASFV and did not react with swine fever virus, porcine reproductive and respiratory syndrome virus, porcine pseudorabies virus and porcine circovirus 2 with good specificity. Three MAbs could recognize ASFV p35 protein, and 15 MAbs recognize p15 protein. Among of them, 14 MAbs heavy chains belong to IgG1 and 4 MAbs heavy chains belong to IgG2a. All light chains were κ. The results of immunohistochemistry showed that 5 MAbs had positive reactivity with the ASFV in lung, tonsil, lymph node of pigs infected with ASFV. The monoclonal antibodies against p62 protein obtained from the study laid a foundation for the establishment of immunological detection method of African swine fever virus and could provide important biomaterials for research on the structure and function of p62 protein.

Key words: African swine fever, p62 protein, monoclonal antibody, immunohistochemistry

CLC Number:

S852.659.1

BAI Chenyu, WANG Tongyan, ZHAO Shaoruo, WANG Heng, HAO Liying, LI Xuefeng, BAI Lulu, DENG Yue, WANG Mengyue, DENG Junhua, LI Xiangdong, ZHANG Guihong, TIAN Kegong. Preparation of Monoclonal Antibodies against Recombinant p62 Protein of African Swine Fever Virus and Its Preliminary Application[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 1074-1082.

References 31

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Preparation of Monoclonal Antibodies against Recombinant p62 Protein of African Swine Fever Virus and Its Preliminary Application

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