两种亲和层析方法纯化兔抗CHO细胞蛋白多克隆抗体的比较

Purification of Rabbit Anti-CHO Cell Protein Polyclonal Antibody by Two Affinity Chromatographic Methods

目的比较两种亲和层析法纯化兔抗CHO细胞蛋白多克隆抗体的效果。方法制备可溶性CHO细胞蛋白,免疫家兔,获得兔免疫血清,采用蛋白A亲和层析和抗原偶联亲和层析两种方法分别纯化兔抗CHO细胞蛋白抗体,ELISA测定抗体效价;SDS-PAGE分析抗体纯度;Western blot分析抗体的特异性。结果兔IgG经蛋白A亲和层析纯化后,ELISA效价约为1∶106,抗体纯度达81.3%;经抗原偶联亲和层析纯化后,ELISA效价可达1∶107,抗体纯度达80.6%。两种方法纯化的抗体特异性均较好。结论抗原偶联亲和层析法更适合纯化兔抗CHO细胞蛋白多克隆抗体。

Objective To compare the purification effects of rabbit anti-CHO cell protein polyclonal antibody by two affinity chromatographic methods.Methods Soluble CHO cell protein was prepared and used for immunization of rabbits to obtain immune sera,based on which rabbit anti-CHO cell protein polyclonal antibody was purified by protein A affinity chromatography and antigen coupled affinity chromatography respectively,then determined for titer by ELISA,and analyzed for purity by SDS-PAGE and for specificity by Western blot.Results The ELISA titer and purity of rabbit IgG after purification by protein affinity chromatography were 1 ∶ 106 and 81.3%,while those after purification by antigen coupled affinity chromatography were 1 ∶ 107 and 80.6%,respectively.Both IgG purified by the two methods showed high specificities.Conclusion As compared with protein A affinity chromatography,antigen coupled affinity chromatography was more suitable for purification of rabbit anti-CHO cell protein polyclonal antibody.