ELISA法筛选适合免疫纯化的单克隆抗体

Screening of Monoclonal Antibody Suitable for Immunopurification Using ELISA Method

目的 筛选适合免疫纯化的单克隆抗体。方法 以乙肝表面抗原 (HBsAg)单克隆抗体和CHO重组细胞表达的HBsAg之ELISA洗脱分析试验为模型 ,建立了一种融合后筛选早期用于选择具有理想免疫纯化结合特性的单克隆抗体。结果 从 3株HBsAg单克隆抗体中 ,利用此法成功地筛选出一株可用于纯化CHO重组细胞表达的HBsAg的单克隆抗体McAb 5D2 。用McAb 5D2 制成的亲和层析柱分离纯化CHO重组细胞表达的HBsAg ,一步纯化倍数可达 342 0倍以上 ,纯度为 96 7% ,收率为 76 %。 结论 本法纯化效果较好 ,且可一次性对多种单克隆抗体及多种洗脱液条件同时检测 ,简便。

Objective Screening of monoclonal antibody suitable for immunopurification.Methods Basing on the principle of elution assay of ELISA of anti HBsAg monoclonal antibodies and HBsAg derived from CHO cells,we have established a simple method to select monoclonal antibodies having dcsired immunopurification Binding properties at the initial stage of screening after a cell fusion .Results Using this method,McAb5D 2,which is applicable for purification of HBsAg expressed by CHO cells,has been successfully screened out from three strains of monoclonal antibcdles.A affinity column prepared by McAb5D 2 is used to purify the HBsAg secreted from the recombinant CHO cells,and the purification factor is up to 3420 folds in one step, the purity and recovery are 96 7%、76% respectively.Conclusion The method is simple, fast and crcdible, it can be used to detect many monoclonal antibodies and various elution condition at one time.