离子交换层析复性重组人γ-干扰素折叠二聚体的形成  

Formation of dimers in refolding of recombinant human interferon-γ by ion-exchange chromatography.

作　　者：靳挺[1] 关怡新[1] 姚善泾[1] 

机构地区：[1]浙江大学化学工程与生物工程学系

出　　处：《浙江大学学报（农业与生命科学版）》2006年第1期101-105,共5页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金资助项目(20476093)

摘　　要：SP Sepharose Fast Flow离子交换层析柱作为蛋白质复性系统,采用尿素梯度法进行重组人γ-干扰素包涵体蛋白质的复性实验.结果表明,尿素梯度离子交换层析法能有效地复性重组人γ-干扰素包涵体,复性后的重组人γ-干扰素的纯度达95%,蛋白收率达54%,比活为7.5×105IU?mg-1.以Superdex 75凝胶作为体积排阻层析介质,对离子交换层析复性的样品进行分析,表明离子交换层析复性的样品中没有重组人γ-干扰素聚集体存在.荧光光谱分析表明重折叠的重组人γ-干扰素的构象接近于其天然二聚体构象.The refolding process was performed by gradually decreasing the concentration of urea in the buffer after the denatured rhIFN-γ proteins had been bound onto the ion-exchange gel SP Sepharose Fast Flow, Results showed that the denatured rhIFN-γ was refolded efficiently by ion-exchange chromatography with a urea gradient and the purity of the refolded rhIFN-γ was up to 95 %. The protein recovery was 54% and specific activity of rhIFN-γ was up to 7.5 ×10^5IU·mg^-1. The chromatogram of size exclusion chromatography with Superdex 75 indicated that the refolded rhIFN-γ didn＇t form any aggregates after renaturation. The conformation of refolded rhIFN-γ was close to the native dimer as shown by fluorescence spectrophotometer characterization.

关 键 词：重组人Γ-干扰素 包涵体 复性 离子交换层析 荧光光谱分析 

分 类 号：Q511[生物学—生物化学]