【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.更多还原

【Abstract】 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 7.5×10⁵ IU·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.更多还原