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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 effects of two kinds of affinity chromatography polyclonal antibodies against rabbit anti-CHO cell protein. Methods The soluble CHO cell protein was prepared and immunized rabbits to obtain rabbit serum. The protein of rabbit anti-CHO cell protein was purified by protein A affinity chromatography and antigen-coupled affinity chromatography. The antibody titer was measured by ELISA. SDS -PAGE analysis of antibody purity; Western blot analysis of antibody specificity. Results After purified by protein A affinity chromatography, the titer of the rabbit IgG was about 1:106 and the purity of the antibody was 81.3%. After purification by antigen-coupled affinity chromatography, the titer of the rabbit IgG was 1:107. The antibody purity Up to 80.6%. Antibodies purified by either method were better specific. Conclusion Antigen-coupled affinity chromatography is more suitable for the purification of rabbit anti-CHO cell protein polyclonal antibody.