鲤春病毒血症病毒单克隆抗体的制备及其特性鉴定  

Preparation and identification of monoclonal antibodies against spring viremia of carp virus

作　　者：张朋[1] 刘荭[2] 陈孝煊[1] 吴志新[1] 余剑敏[1] 杜娟[1] 兰文升[2] 史秀杰 郑晓聪[2] 何俊强[2] 贾鹏[2] 申斯思[2] 朱家增[2] 

机构地区：[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

出　　处：《中国预防兽医学报》2011年第4期305-308,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家863项目(2006AA100306);国家质检总局科研项目(2007IK022);深圳出入境检验检疫局科研项目(SZ2007105)

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∶160000～1∶640000。亚型鉴定结果表明,这些MAb分属2个亚型(1F1、3E1,IgG2a;3F5、4F9,IgG1),轻链均为Κ链。Western blot分析显示,MAb1F1、3F5、4F9能特异性地识别SVCV的N蛋白(47ku),3E1能特异性地识别SVCV的G蛋白(69ku)。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。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 ku of glycoprotein. Epitope reaction analysis demonstrated that 1F1, 3F5, and 4F9 might recognize the same epitope, while 3E1 was different. Indirect immunofluorescence assay showed that all of these MAbs reacted positively with the virus in the SVCV-infected cells. These antibodies will be useful tools to establish immunological detection methods of SVCV.

关 键 词：鲤春病毒血症病毒 单克隆抗体 N蛋白 

分 类 号：S852.65[农业科学—基础兽医学]