玻勒虹彩病毒单克隆抗体的制备及初步应用  

Preparation and preliminary application of monoclonal antibodies against Bohle virus

作　　者：景宏丽[1] 林祥梅[1] 梅琳[1] 张旻[1] 王娜[1] 高隆英 张利峰[3] 吴绍强[1] 

机构地区：[1]中国检验检疫科学研究院,北京100029 [2]深圳蛇口出入境检验检疫局,广东深圳518067 [3]北京出入境检验检疫局,北京100026

出　　处：《大连海洋大学学报》2015年第5期505-508,共4页Journal of Dalian Ocean University

基　　金："十二五"国家科技支撑计划项目(2013BAD12B02);质检公益行业科研专项(201310221)

摘　　要：为了制备抗玻勒虹彩病毒(Bohle virus,BIV)的单克隆抗体,应用杂交瘤细胞技术,制备了1株稳定分泌抗玻勒虹彩病毒单克隆抗体的杂交瘤细胞株(3A4),并对其单克隆抗体特性进行分析和初步应用。结果表明:单抗3A4腹水效价为105以上,属于Ig G3型,κ链;单抗3A4与大鲵虹彩病毒(ADIV)、流行性造血器官坏死病毒(EHNV)发生反应,与其他病毒株和细胞株不发生反应;单抗3A4可识别相对分子质量为50 000的蛋白;应用捕获ELISA方法对样品进行检测,建立的捕获ELISA方法能用于蛙病毒的检测。该单抗为BIV的快速诊断及流行病学监测提供了检测试剂。One monoclonal antibody （ MAb ） （ 3 A4 ） against Bohle virus （ BIV ） was prepared by immunizing BALB/c mice and the MAb （3A4） characteristics and application were studied by indirect enzyme-linked immuno-assay （ ELISA） and western-blotting. An antigen-capture ELISA was developed and the samples were detected by BIV. The indirect ELISA revealed that the titer of MAb （3A4） ascites was 105 , and belonged to IgG3 subclass andκ chain. The MAb （3A4）did not react with other fish viruses or fish cells lines in the ELISA test except BIV, An-drias davidianus iridovirus （ ADIV） and epizootic haematopoietic necrosis virus （ EHNV） . In the western blotting using MAb 3A4, a 50 000 protein band was observed when BIV was used as antigen. The antigen-capture ELISA confirmed that the ELISA was able to detect virus from amphibian, and the obtained MAb was applied to rapid di-agnosis and detection of BIV in aquaculture.

关 键 词：玻勒虹彩病毒(BIV) 捕获ELISA 单克隆抗体 

分 类 号：S941[农业科学—水产养殖]