柱层析法纯化基因工程HBsAg中试工艺的研究

由乙肝病毒S基因转化的哺乳动物细胞培养收液，经过Butyl-S-SepharoseFF层析，DEAESepharoseFF层析和Sepharose4FF层析，可获HBsAg纯品。产品检定结果表明，PAGE和SDS-PAGE电泳纯度均合格，小牛血清残余量、细胞DNA残余量和动物免疫效力也均符合规定标准．HBsAg终收率达40％左右．在此工艺中，疏水作用柱层析的纯化效率比较高，可去除绝大部分杂蛋白和细胞DNA。

Pilot Study on Purification Process of Recombinant HBsAg by Column Chromatography

After HBsAg S gene transformed CHO cell, culture fluid was purified by ButylS-Sepharose FF, DEAE Sepharose FF and Sepharose 4 FF chromatography, and purified HBsAg was obtained. PAGE and SDS-PAGE proved that the averaged purity of the purified HBsAg reached 96 %. The residual calf serum protein and host cell DNA content of the purified products also consistent with the standards. Animal test showed that the immunogenicity of it was better than that of plasma derived vaccine. The total recovery of HBsAg was about 40%. The hydrophobic column chromatography used for purification is highly efficient, and the majority of protein impurities and host cell DNA can be removed.