基因工程β型干扰素的分离纯化  

Purification of recombinant human fibroblast interferon produced in Escherichia coli

作　　者：白东亭[1] 许丽锋[2] 郑秋君[2] 陈滨[2] 蒋盘宏[2] 赵睿[3] 刘国诠[3] 吴淑华[4] 

机构地区：[1]中国药品生物制品检定所,北京100050 [2]卫生部北京生物制品研究所 [3]中国科学院化学研究所 [4]中国预防医学科学院病毒学研究所

出　　处：《中华微生物学和免疫学杂志》1996年第6期452-456,共5页Chinese Journal of Microbiology and Immunology

摘　　要：人β型干扰素工程菌经发酵培养、裂解后，用硫酸铵分级分离及有机溶剂抽提－酸沉淀两种方法对粗制品进行部分纯化，然后用疏水层析、凝胶过滤和旋转等电聚焦电泳对部分纯化品再纯化，结果显示：部分纯化的基因工程人β型干扰素经疏水层析、凝胶过滤后，有较高的回收率（胶者为９３．３６％，后者为８０．６％）；经旋转等电聚焦电泳，一步纯化３１．７５倍，回收率为５６．２５％，比活性达１．０２×１０＾７Ｕ／ｍｇ蛋白。The rhIFN-p, contained in the suspension of disrupted engineered bacteria , was purified paratially by saturated ammonium sulphate or organic extract-acid precipitation method , then it was purified by chromatography of CPG-PEG in HPLC ,5ephacryl S-200 and Rotofor-IEF. The results showed that CPIE-PEC and S-200 had less effect on activity of rhIFN-β(i. e , high recovery rate , separately 93. 36％ and 80. 6％ ) ,but purification efficiency was lower than that of Rotofor-IEF, rhIFN-β could be partially purified by Rotofor-IFiF by 3l. 75 times with a recovery rate of 56. 25 ％ and the specific activity of 1. 02 X 107U/ml protein.

关 键 词：基因工程 β型干扰素 疏水层析 旋转等电聚焦 

分 类 号：Q78[生物学—分子生物学] TQ929.1[轻工技术与工程—发酵工程]