中国兽医科学2024，Vol.54Issue(8)：1065-1071,7.DOI:10.16656/j.issn.1673-4696.2024.0139
猪伪狂犬病病毒gB蛋白单克隆抗体的制备及鉴定
Preparation and identification of a monoclonal antibody against pseudorabies virus gB protein
摘要
Abstract
In order to prepare monoclonal antibodies(MAbs)against the gB protein of porcine pseu-dorabies virus(PRV),the main antigen region(540-734aa)of gB protein was expressed by a prokaryotic expression system,and the recombinant protein was used to immunize BALB/c mice.Using hybridoma tech-nology,a MAb against the gB protein was obtained through the cell screening and subcloning,and named as 1C10.Indirect immunofluorescence assay(IFA)and Western-blot assay verified that 1C10 has a good specificity and could recognize the gB protein expressed in cells infected with PRV.The identification of isotyping revealed that the 1C10 was of IgG2a type for heavy chain and kappa type for light chain.The ELISA data showed that the purified ascites was higher than 1:128 000.In summary,PRV gB protein was successfully expressed and a MAb against PRV gB protein were prepared in this study,laying the founda-tion for further study on the structure and function of PRV gB protein,as well as the establishment of diagnostic methods and pathogenic mechanisms in the later stage.关键词
猪伪狂犬病病毒/gB蛋白/原核表达/单克隆抗体Key words
porcine pseudorabies virus/gB protein/prokaryotic expression/monoclonal antibodies分类
农业科技引用本文复制引用
马国祥,袁聪,施磊,郭庆勇,翟少华,郑海学,王琦,曹丽艳,张娟,张宇,万颖,孔祥雨,李想通,王娅婷,段月月..猪伪狂犬病病毒gB蛋白单克隆抗体的制备及鉴定[J].中国兽医科学,2024,54(8):1065-1071,7.基金项目
四川省科技计划重点研发项目(2022YFN0008) （2022YFN0008）
四川省青年学者基金项目(21JCQN0175) （21JCQN0175）
中国农业科学院创新工程项目(ASTIP2022-34-IUA-07) （ASTIP2022-34-IUA-07）
中国农业科学院青年英才项目(21ZD3NA001) （21ZD3NA001）
