疏水作用层析在重组乙肝表面抗原中的应用研究.pdfVIP

疏水作用层析在重组乙肝表面抗原中的应用 周卫斌 闭静秀 李岩 张焱 黄永东 苏志国∗∗ （中国科学院过程工程研究所生化工程国家重点实验室，北京 100080 ） 摘 要 实际生产中重组CHO- 乙肝表面抗原的分离纯化效果不佳，为提高活性回收率和纯化效果，降低 生产成本，本文对该疫苗分离纯化工艺中影响疏水作用层析过程的因素进行了考察。除疏水层析介质、pH 、 硫酸铵浓度等条件外，样品浓度、进料量、层析柱高度及高径比、样品预处理乃至温度等均是影响重组乙 肝表面抗原分离纯化效果的因素。其中，用硫酸铵沉淀预处理细胞培养上清液样品对疏水层析影响很大， 经硫酸铵沉淀处理后的细胞培养上清液再经疏水层析处理可将进料量由原来的样品体积与柱体积之比由原 来的10:1 左右提高到为20～25 ：1，并可在提高原活性回收率的基础上将纯化倍数提高2～3 倍。本实验进 一步证实通过优化疏水层析过程降低CHO-HBsAg 分离纯化工艺成本的现实可行性。 关键词 重组乙肝表面抗原，分离纯化，疏水作用层析 HYDROPHOBIC INTERACTION CHROMATOGRAPHY IN ISOLATION AND PURIFICATION OF RECOMBINANT HEPATITIS B SURFACE ANTIGEN ∗ Weibin Zhou, Jingxiu Bi, Yan Li, Yan Zhang, Yongdong Huang, Zhiguo Su (National Laboratory of Biochemical Engineering, Chinese Academy of Sciences, Beijing 100080, China) Abstract: In order to improve the recovery and purification(fold) of the commercialized recombinant CHO-HBsAg, the condition of hydrophobic interaction chromatography was modified. In addition to kinds of gel, pH and concentration of ammonium sulfate, many parameters contributed to effect of purification, such as samples, ratio of sample volume to column volume, column dimension, temperature, etc. Sample-pretreatment with ammonium sulfate precipitation (ASP) acted a very important part. The HIC of sample pretreated with ASP had higher recovery of HBsAg and 2~3 times higher purification(fold), and the capacity was improved from ratio of sample volume to column volume with value 10:1 to 20~25:1. Moreover, lower cost was obtained in the modified process of HIC in purification of HBsAg. Keywords: recombinant HBsAg, isolation and purification, hydrophobic interaction chromatography 引 言 乙型肝炎是危胁我国人民健康的大敌。据不完全统计，我国人口至少有10% 为乙肝病毒携带者[1]。基因工程乙肝疫苗是迄今为止最好的防治该疾病的方法[2, ∗通讯联系人：苏志国，+86-10 E-mail: zgsu@home.ipe.ac.cn 国家高科技发展计划（863 计划）的基金资助（No. 2002AA217031 ）