鹿流行性出血病病毒单克隆抗体的制备与鉴定  

Preparation and Characterization of Monoclonal Antibodies Against Epizootic hemorrhagic disease virus

作　　者：郭莹洁[1,2] 杨俊兴[1] 花群义[1] 徐聪[1,2] 林庆燕[1] 阮周曦[1] 陈兵[1] 卢体康[1] 詹爱军[2] 

机构地区：[1]深圳出入境检验检疫局动植物检验检疫技术中心．广东深圳518001 [2]云南农业大学动科院,云南昆明650201

出　　处：《动物医学进展》2009年第3期14-17,共4页Progress In Veterinary Medicine

基　　金：国家863计划项目(2006AA10Z445)

摘　　要：为了更好的研究鹿流行性出血病病毒(EHDV)的生物学特性和建立EHDV免疫学检测方法,本研究用纯化的EHDV免疫Balb/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞融合,经间接ELISA筛选,有限稀释法克隆,获得了2株(1A5和8H6)能够稳定分泌抗EHDV单抗的杂交瘤细胞株。其细胞培养上清效价分别为1∶256和1∶512,腹水效价分别为1∶1024000和1∶2048000;McAb 1 A5和8H6的相对亲和力分别为0.9mg/L和0.7mg/L。间接ELISA结果显示,2株单抗仅与EHDV反应,不与蓝舌病病毒、赤羽病病毒、水疱性口炎病毒、小反刍兽疫病毒和BHK细胞反应,表明2株单抗特异性良好。这2株单抗的获得为研究EHDV生物学特性和建立EHDV检测方法奠定了基础。For studying of the biological characteristics and establishing immunological test method of Epizootic hemorrhagic disease virus （EHDV）, Balb/c mice, 6-8 weeks old, were immunized with purified EHDV. Monoclonal antibodies （McAbs） 1A5 and 8H6 against EHDV were produced by fusing SP2/0 myeloma cells with spleen cells of Balb/c mice. The indirect ELISA results showed that the titers of McAbs in hybridoma cell supernatant and ascites werel ： 256, 1 ： 512 and 1 ： 1 024 000, 1 ： 2 048 000,respectively. The relative affinity of the McAb 1A5 and 8H6 were 0.9 mg/L and 0.7 mg/L. Both MeAbs reacted with EHDV but not with bluetongue virus, Akabane disease virus, vesicular stomatitis virus and Peste des petits ruminants virus,as determined by ELISA. The McAbs are useful tools for studying biological characteristics and developing diagnostic techniques of EHDV.

关 键 词：鹿流行性出血病病毒 单克隆抗体 制备 

分 类 号：S852.659.4[农业科学—基础兽医学]