摘要
为了获得我国兔出血症病毒2型(RHDV2)SC株特异性单克隆抗体,研究以纯化RHDV2作为抗原免疫BALB/c小鼠,4次免疫后取脾细胞与SP2/0细胞融合,经间接ELISA方法筛选,获得一株能够稳定分泌RHDV2 VP60蛋白特异性单克隆抗体的杂交瘤细胞,命名为6B3。经Western blot和间接免疫荧光试验验证,该单克隆抗体可识别RHDV2,不与RHDV1反应,具有良好的毒株特异性。亚型鉴定结果为IgG3,轻链为Kappa型,腹水ELISA效价为1∶80 000。本研究得到的针对RHDV2的特异性单抗,将对我国兔出血症的诊断及流行病学调查提供有效制剂。
In this study,BALB/c mice were immunized with purified RHDV2 as antigen to obtain specific monoclonal antibody against the VP60 protein of rabbit hemorrhagic disease virus type 2 SC strain.After 4 times of immunization,spleen cells were collected and fused with SP2/0 cells.After multiple indirect ELISA methods,a hybridoma cell named 6B3 that could secrete RHDV2-specific monoclonal antibody stably was obtained.Western blot and indirect immunofluorescence assay confirmed that the monoclonal antibody reacted with RHDV2,but not with RHDV1.The titer of the monoclonal antibody was up to 1:80000.The subtype identification result showed that 6B3 was IgG3 and of the Kappa light chain.The specific monoclonal antibody against RHDV2 obtained in this study would be great significance for the diagnosis and epidemiological investigation of rabbit hemorrhagic disease in China.
作者
饶华琴
魏后军
仇汝龙
陈萌萌
胡波
范志宇
刘晓娜
薛家宾
宋艳华
钱莺娟
王芳
RAO Huaqin;WEI Houjun;QIU Rulong;CHEN Mengmeng;HU Bo;FAN Zhiyu;LIU Xiaona;XUE Jiabin;SONG Yanhua;QIAN Yingjuan;WANG Fang(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences/Key Laboratory for Veterinary Bio-Product Engineering,Ministry of Agriculture and Rural Affairs,Nanjing 210014,China)
出处
《畜牧与兽医》
CAS
北大核心
2023年第7期58-63,共6页
Animal Husbandry & Veterinary Medicine
基金
江苏省农业科技自主创新项目[CX(20)3158]
现代农业产业技术体系建设专项资金资助项目(CARS-43-C-1)。
关键词
兔出血症病毒2型
VP60蛋白
单克隆抗体
毒株特异性
鉴定
rabbit hemorrhagic disease virus type 2
VP60 protein
monoclonal antibody
strain specificity
identification
