PEG沉淀结合层析分离重组乙肝病毒表面抗原  

Purification of Recombinant Hepatitis B Virus Surface Antigen with PEG Precipitation and Chromatography

作　　者：张焱[1] 闭静秀[1] 周卫斌[1] 李岩[1] 董爱华 孙娟[1,3] 苏志国[1] 

机构地区：[1]中国科学院过程工程研究所生化工程国家重点实验室 [2]华北制药集团新药中心 [3]北京科技大学土木与环境学院,北京100083

出　　处：《过程工程学报》2006年第1期100-103,共4页The Chinese Journal of Process Engineering

基　　金：国家863计划资助项目(编号:2002AA217031)

摘　　要：应用聚乙二醇(PEG)沉淀结合层析技术纯化乙肝病毒表面抗原(HBsAg),对PEG浓度、pH值、离子强度等影响PEG沉淀的因素进行了正交实验研究.结果表明,浓度为0.12g/mL的PEG6000在4℃,pH9.0条件下沉淀HBsAg纯化效果比较理想,纯化倍数达3.7,回收率96.8%,有效地去除了生物大分子杂质和牛血清白蛋白(BSA),PEG在后续的凝胶过滤中可以除去,整个工艺的回收率可提高到41%.Chromatography integration is being widely used in the purification of hepatitis B virus surface antigen （HBsAg） expressed by recombinant Chinese hamster ovary cell line. The low final product recovery rate limits manufactory production and HBsAg vaccine application. Polyethylene glycol （PEG） precipitation is applied to improve HBsAg yield. The effects of PEG concentration, pH value and ionic intensity on PEG precipitation were investigated by orthogonal design of the experiments. Under the optimized conditions of 0.12 g/mL PEG6000, 4 ℃, pH 9.0, the recovery rate of HBsAg was up to 96.8% while the purification factor was 3.7. Furthermore, the macromolecular impurity and bovine serum albumin were effectively removed by PEG precipitation. The total recovery rate of HBsAg was improved to 41% and PEG could be removed by gel filtration chromatography in the final product.

关 键 词：重组乙肝病毒表面抗原 PEG沉淀 分离纯化 层析技术 

分 类 号：Q819[生物学—生物工程]