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作 者:徐聪[1,2] 杨俊兴[1] 花群义[1] 阮周曦[1] 陶虹[1] 段纲[2] 郭莹洁[1,2] 陈兵[1] 詹爱军[1]
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]云南农业大学动物科学技术学院,云南昆明650201
出 处:《动物医学进展》2009年第6期1-4,共4页Progress In Veterinary Medicine
基 金:国家863计划项目(2006AA10Z445)
摘 要:用纯化的蓝舌病病毒(BTV)免疫Balb/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合,经间接ELISA方法筛选,有限稀释法克隆,获得2株稳定分泌抗BTV特异性单克隆抗体的杂交瘤细胞株(1F5和4E5)。其细胞培养上清ELISA效价分别为1∶512和1∶256,腹水ELISA效价分别为1∶512 000和1∶128 000。亚型鉴定表明,1F5和4E5分别为IgG1和IgG2a。ELISA结果显示,2株单克隆抗体仅与BTV反应,不与其他相关病毒反应,表明2株单克隆抗体特异性良好。2株单克隆抗体1F5和4E5的相对亲和力指数分别为5.14×106mol/L和6.71×106mol/L。这2株单克隆抗体的获得为建立BTV免疫学检测方法奠定了基础。Monoclonal antibodies (McAbs) 1F5 and 4E5 against bluetongue virus (BTV) were produced by fusing SP2/0 myeloma cells with spleen cells of Balb/c mice immunized with purified BTV,and selected by indirect enzyme linked immunosorbent assay (ELISA). The ELISA titers of antibodies in supernatants were 1:512 and 1 :256, respectively. The titers of antibodies in mouse ascites were 1: 512 000 and 1:128 000,respectively. Antibody subclasses analysis showed that McAb 1F5 and MCAb 4E5 were IgG1 and IgG2a,respectively. The relative affinity indexes of the McAbs were 5.14×10^6 mol/L and 6.71×10^6 mol/L. The result of ELISA showed that both MeAhs specifically reacted to BTV but not with other viruses. The McAbs are useful tools for developing diagnostic techniques of BTV.
分 类 号:S852.659.4[农业科学—基础兽医学]
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