新城疫病毒单克隆抗体的制备及ELISA检测方法的建立  

Development of monoclonal antibody against NDV and establishment of ELISA detection method

作　　者：赵坤[1] 李任峰[1] 赵朴[1] 赵恒章[1] 常新耀[1] 

机构地区：[1]河南科技学院动物科学学院,河南新乡453003

出　　处：《河南农业大学学报》2011年第4期428-431,共4页Journal of Henan Agricultural University

基　　金：河南省基础与前沿技术研究计划项目(072300430020)

摘　　要：将复性的重组NDVHN蛋白免疫BALB/c小鼠的脾细胞与SP2/0骨髓瘤细胞融合制备杂交瘤.通过ELISA筛选,获得了1株稳定分泌抗HN单克隆抗体(MAb)的杂交瘤细胞4E11.ELISA和Western-blot结果表明,该MAb能与NDV特异性反应,而不与H9亚型AIV,IBDV,EDS76,IBV和CIAV交叉反应.利用该MAb和抗NDV多克隆抗体IgG建立了双抗体夹心ELISA.试验结果表明该DAS-ELISA检测NDV的敏感性可达8个血凝单位.BALB/c mice were immunized with the recombinant HN protein of NDV.Hybridomas was generated by the fusion of the spleenocytes from these mice with SP2/0 myeloma cells.Hybridoma cells were screened by ELISA.One hybridoma cell line（4E11） stable in secreting anti-HN monoclonal antibodies（MAb） was generated.Specificity of the MAbs was identified by ELISA and Western blot.The results showed that the MAb can specifically react with NDV without cross-reactivity to H9-AIV,IBDV,EDS76,IBV and CIAV.Double antibody sandwich ELISA（DAS-ELISA）using the MAb and anti-NDV IgG polyclonal antibody was developed.Experimental results proved that 8 hemagglutination units（HAU）of NDV can be detected by the DAS-ELISA.It is helpful for promoting detection assay and pathogenesis studies on NDV.

关 键 词：新城疫病毒 单克隆抗体 ELISA 

分 类 号：S852[农业科学—基础兽医学]