离子交换层析分离重组乙肝病毒表面抗原过程中的介质筛选及条件优化研究.pdfVIP

离子交换层析分离重组乙肝病毒表面抗原过程中的 介质筛选及条件优化 黄永东，闭静秀，周卫斌，李岩，张焱，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 探索汉逊酵母表达的重组乙肝病毒表面抗原HBsAg （y-HBsAg ）在离子交换介质上的分离。首先考 察GE Healthcare 公司生产的DEAE-Sepharose FF 用于y-HBsAg 分离纯化的可行性，并优化了其层析条件， 同时考察了采用本课题组所建立的包括离子交换层析在内的分离纯化工艺制备y-HBsAg 纯品的可行性。此 外，还考察了自制介质DEAE-Sepharose 6FF 用于y-HBsAg 分离纯化的可行性。结果表明，两种介质的性 能相当，单步的HBsAg 回收率和纯化倍数分别可以达到60%和3 左右，包括离子交换层析在内的分离纯化 工艺能够以20%左右的HBsAg 回收率得到HBsAg 纯品。此外，自制介质DEAE-Sepharose 6FF 也可用于中 国仓鼠卵巢细胞表达的HBsAg （CHO-HBsAg ）的分离纯化，应用前景广阔。 关键词 离子交换层析 汉逊酵母 乙肝病毒表面抗原 中图分类号 R392 －33 文献标识码 A SCREENING THE MEDIA AND OPTIMIZATION THE OPERATION OF ION-EXCHANGE CHROMATOGRAPHY FOR THE PURIFICATION OF RECOMBINANT HEPATITIS B VIRUS SURFACE ANTIGEN HUANG Yongdong, BI Jingxiu, ZHOU Weibin, LI Yan, ZHANG Yan and SU Zhiguo1∗ (National Laboratory of Biochemical Engineering, Chinese Academy of Sciences, Beijing 100080, China) Abstract The ion-exchange chromatography (IEC) purification process of recombinant Hepatitis B virus antigen (r-HBsAg) derived from Hansenula polymorpha yeast (y-HBsAg) system was explored. Firstly, the ion-exchange media DEAE-Sepharose FF from GE Healthcare was used to purify y-HBsAg, then the chromatographic condition was optimized. Additional, a downstream process including IEC, established by our research group was carried out to prepare y-HBsAg. Further, the feasibility of separating y-HBsAg with homemade anion ion-exchange media of DEAE-Sepharose 6FF was studied. The results demonstrated that about 60% HBsAg recovery and 3 of purification fold (PF) were obtained during the process of IEC, and the purified y-HBsAg can be obtained with HBsAg recovery about 20% using the new downstream process established. The homemade DEAE-Sepharose 6FF has the same chromatographic efficiency as that of DEAE-Sepharose FF from GE Healthcare. Moreov