番茄花叶病毒单克隆抗体的制备及检测应用

Production of Monoclonal Antibodies to Tomato Mosaic Virus and Application in Virus Detection

用番茄花叶病毒 (ToMV)免疫的BALB c鼠脾细胞与SP2 0鼠骨髓瘤细胞融合 ,经筛选克隆 ,获得 4株能稳定传代并分泌抗ToMV单克隆抗体 (MAb)的杂交瘤细胞 ,其中 2株能同时检测ToMV和烟草花叶病毒 (TMV) ,各单克隆抗体腹水ELLSA效价在 1∶3 2 0 0 0～ 1∶1 0 2 4 0 0 0之间。经TAS ELISA测定 ,4株单克隆抗体检测病汁液的稀释度均能达到 1∶2 0 0 0倍以上。 4株单克隆抗体与其他病毒无交叉反应。Western blot分析表明 ,其中两株与ToMV1 7 6kD的外壳蛋白亚基有特异反应 ,而另两株无反应 ,推测它们是针对构象决定簇的抗体。

Four hybridoma cell lines secreting monoclonal antibodies(MAbs) against Tomato mosaic virus(ToMV) were produced by fusing mouse myeloma cells(SP2/0) with spleen cells from BAL B/cimmunized by the ToMV particle.The four MAbs could specifically react with ToMV,and the MAbs from two cell lines can react with ToMV and TMV simultaneously.The titres of ascitic fluids of four MAbs ranged from 1∶32 000 to 1∶102 400 0 with ELISA,and the sensitivity for detection virus from the plant sap reached over 1∶2000 dilution.The MAbs didn't crossreact with other plant viruses.The result of Western blot showed that two MAbs can react with the 17.6 kD ToMV coat protein submit specifically,while the other two MAbs can not react with it,they are supposed to against conformational determinants of ToMV CP.