疏水作用色谱法同时纯化及复性基因重组人干扰素-α

 使用高效疏水作用色谱直接从大肠杆菌表达的基因重组人干扰素 α(rhIFN α)包涵体的裂解液中纯化了rhIFN α ,并在纯化的同时获得了高的复性效率 ,使复性和纯化一步完成 ,大大地简化了操作步骤。用凝胶排阻色谱对该法纯化的rhIFN α进行了纯度测定 ,纯度达到 95 %以上。该法的活性回收率分别比稀释法和透析法高 1 0倍和 1 6倍。

Simultaneous Purification and Renaturation of Recombinant Human Interferon-Alpha Expreessed by E.coli by High-Performance Hydrophobic Interaction Chromatography

One-step simultaneous purification and renaturation of interferon-alpha from the inclusion body of E. coli expreessed by high-performance hydrophobic interaction chromatography(HPHIC) are presented. The inclusion body was dissolved by 8 mol/L guanidine hydrochloride. The sample lysis was centrifuged and the solution was renaturated by three different methods, i.e. hydrophobic interaction chromatography, dilution and dialysis. The total activity yield renaturation by hydrophobic interaction chromatography was 2 times and 2.6 times as much as that by dilution and by dialysis. The purity of the target product was 95% according to high-performance size exclusion chromatography and the specific activity was 1.8 x 10(8) IU/mg. The method developed is simpler and more efficient than others.