采用疏水相互作用色谱(HIC)对还原变性核糖核酸酶A (RNase A)在疏水性液-固界面上的复性进行了研究.详细讨论了流动相中脲的浓度、还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)的比例、流动相pH和变性蛋白质浓度对还原变性RNase A复性效率和质量回收率的影响.结果表明,在最优化的复性条件(流动相中含有2.0 mol/L 脲,GSH/GSSG的浓度比为8∶1,流动相pH为8.0)下,还原变性RNase A能完全复性.当变性蛋白质质量浓度为5.0 mg/mL 时,还原脲变性RNase A的活性回收率和质量回收率分别为98.0%和61.9% ,还原胍变性RNase A分别为100.1%和66.8% .研究表明HIC是还原变性蛋白质复性的有力工具之一,可为蛋白质复性研究提供新方法和新思路.
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