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鲤春病毒血症病毒单克隆抗体的制备及其特性鉴定
引用本文:张朋,刘荭,陈孝煊,吴志新,余剑敏,杜娟,兰文升,史秀杰,郑晓聪,何俊强,贾鹏,申斯思,朱家增.鲤春病毒血症病毒单克隆抗体的制备及其特性鉴定[J].中国预防兽医学报,2011,33(4).
作者姓名:张朋  刘荭  陈孝煊  吴志新  余剑敏  杜娟  兰文升  史秀杰  郑晓聪  何俊强  贾鹏  申斯思  朱家增
作者单位:1. 华中农业大学水产学院,湖北武汉,430070
2. 深圳出入境检验检疫局,广东深圳,518045
基金项目:国家863项目,国家质检总局科研项目,深圳出入境检验检疫局科研项目
摘    要:为获得针对鲤春病毒血症病毒(SVCV)特异性的单克隆抗体(MAb),以纯化的SVCV为抗原,免疫BALB/c小鼠.将免疫鼠的脾细胞与SP2/0骨髓瘤细胞融合,采用间接ELISA法筛选获得4个能稳定分泌抗SVCV MAb的杂交瘤细胞株;4个杂交瘤细胞制备腹水的MAb效价为1:160 000~1:640 000.亚型鉴定结果表明,这些MAb分属2个亚型(1F1、3E1,IgG2a;3F5、4F9,IgGl),轻链均为K链.Western blot分析显示,MAb1F1、3F5、4F9能特异性地识别SVCV的N蛋白(47 ku),3E1能特异性地识别SVCV的G蛋白(69 ku).采用相加ELISA法对抗原表位分析结果显示,1F1、3F5、4F9可能识别相同的表位,3E1则识别不同的表位.间接免疫荧光试验结果显示4株MAb均能对染毒病灶产生特异性的荧光染色.这些MAb的制备为SVCV免疫学检测方法的建立奠定了基础.

关 键 词:鲤春病毒血症病毒  单克隆抗体  N蛋白

Preparation and identification of monoclonal antibodies against spring viremia of carp virus
ZHANG Peng,LIU Hong,CHEN Xiao-xuan,WU Zhi-xin,YU Jian-min,DU Juan,LAN Wen-sheng,SHI Xiu-jie,ZHENG Xiao-cong,HE Jun-qiang,JIA Peng,SHEN Si-si,ZHU Jia-zeng.Preparation and identification of monoclonal antibodies against spring viremia of carp virus[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(4).
Authors:ZHANG Peng  LIU Hong  CHEN Xiao-xuan  WU Zhi-xin  YU Jian-min  DU Juan  LAN Wen-sheng  SHI Xiu-jie  ZHENG Xiao-cong  HE Jun-qiang  JIA Peng  SHEN Si-si  ZHU Jia-zeng
Institution:ZHANG Peng1,LIU Hong2*,CHEN Xiao-xuan1*,WU Zhi-xin1,YU Jian-min1,DU Juan1,LAN Wen-sheng2,SHI Xiu-jie2,ZHENG Xiao-cong2,HE Jun-qiang2,JIA Peng2,SHEN Si-si2,ZHU Jia-zeng2 (1. College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China,2. Shenzhen Entry & Exit Inspection and Quarantine Bureau,Shenzhen 518045,China)
Abstract:To prepare specific monoclonal antibodies (MAbs) against spring viremia of carp virus (SVCV), mouse myeloma cells (SP2/0) were fused with spleen cells from BALB/c immunized with purified SVCV. Four hybridoma cell lines secreting MAbs against SVCV were obtained by indirect ELISA. The titres of ascitic fluids of the MAbs ranged from 1:160,000 to 1:640,000. Western blot analysis revealed that three MAbs (1F1, 3F5 and 4F9) specifically recognized to 47 ku of nucleoprotein, while the MAb 3E1 only reacted with 69...
Keywords:spring viremia of carp virus  monoclonal antibodies  N protein  
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