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›› 2006, Vol. 6 ›› Issue (1): 100-103.

• 生化工程专栏 • 上一篇    下一篇

PEG沉淀结合层析分离重组乙肝病毒表面抗原

张焱,闭静秀,周卫斌,李岩,董爱华,孙娟,苏志国   

  1. 南京工业大学材料科学与工程学院
  • 出版日期:2006-02-20 发布日期:2006-02-20

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

ZHANG Yan,BI Jing-xiu,ZHOU Wei-bin,LI Yan,DONG Ai-hua,SUN Juan,SU Zhi-guo   

  1. College of Materials Science & Technology, Nanjing university of Technology
  • Online:2006-02-20 Published:2006-02-20

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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0.12 g/mL的PEG6000在4℃, pH 9.0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%.

关键词: 重组乙肝病毒表面抗原, PEG沉淀, 分离纯化, 层析技术

Abstract: 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.

Key words: recombinant hepatitis B virus surface antigen, PEG precipitation, purification, chromatography