基因工程β型干扰素的分离纯化

Purification of recombinant human fibroblast interferon produced in Escherichia coli

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.