中国动物传染病学报2023，Vol.31Issue(5)：188-193,6.
非洲猪瘟病毒p72蛋白单克隆抗体的制备及鉴定
Preparation and Characterization of Monoclonal Antibodies Against p72 Protein of African Swine Fever Virus
摘要
Abstract
In order to obtain monoclonal antibodies(MAbs)specific for the p72 eukaryotic protein of African swine fever virus(ASFV)by hybridoma technique,the recombinant plasmid pCMV-p72 and B602L plasmid were co-transfected into HEK293F cells via transient transfer method for the expression and further purification of p72-His recombinant protein.Then,the purity was analyzed by SDS-PAGE and indirect ELISA.The BALB/c mice were immunized with the purified recombinant p72-His protein and anti-p72 MAbs were prepared by cell fusion and screening.After rounds of selection,four hybridoma cell lines(2B2-1,34C10,22D3 and 32H5)were found to secret MAbs against ASFV p72 protein.The isotypes of these MAbs were identified as IgG1/κ or IgG2b/κ.The ascitic titers of these MAbs were higher than 6.4×104.The results of immunofluorescence showed that these 4 MAbs specifically recognized ASFV p72 protein.Further testing found that the affinity of these MAbs was 22D3>32H5>2B2-1 and34C10.Epitope competition test also revealed that these MAbs recognized different antigen binding sites.Additionally,2B2-1 and 32H5 were a good antibody pair for detection of ASFV while 34c10 was a competitor.In summary,these four hybridoma cell lines stably secreted MAbs against p72 protein with high specificity and affinity,which was beneficial to further research and development of ASFV serological diagnosis methods.关键词
非洲猪瘟病毒/p72蛋白/真核表达/单克隆抗体Key words
African swine fever virus/protein p72/Eukaryotic expression/monoclonal antibody分类
农业科技引用本文复制引用
王彩霞,冯春燕,于浩洋,宋晓晖,仇松寅,刘晓飞,吴绍强,林祥梅..非洲猪瘟病毒p72蛋白单克隆抗体的制备及鉴定[J].中国动物传染病学报,2023,31(5):188-193,6.基金项目
十三五国家重点研发计划项目(2016YFD0501100) （2016YFD0501100）
中国检验检疫科学研究院基本科研业务经费(2020JK020) （2020JK020）
