Jin Ting, Guan Yi-Xin, Yao Shan-Jing, Lin Dong-Qiang, Cho Man-Gi
Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China.
Biotechnol Bioeng. 2006 Mar 5;93(4):755-60. doi: 10.1002/bit.20763.
A refolding strategy was described for on-column refolding of recombinant human interferon-gamma (rhIFN-gamma) inclusion bodies by expanded bed adsorption (EBA) chromatography. After the denatured rhIFN-gamma protein bound onto the cation exchanger of STREAMLINE SP, the refolding process was performed in expanded bed by gradually decreasing the concentration of urea in the buffer and the refolded rhIFN-gamma protein was recovered by the elution in packed bed mode. It was demonstrated that the denatured rhIFN-gamma protein could be efficiently refolded by this method with high yield. Under appropriate experimental conditions, the protein yield and specific activity of rhIFN-gamma was up to 52.7% and 8.18 x 10(6) IU/mg, respectively.
描述了一种通过扩张床吸附（EBA）色谱法对重组人干扰素-γ（rhIFN-γ）包涵体进行柱上重折叠的策略。变性的rhIFN-γ蛋白结合到STREAMLINE SP阳离子交换剂上后，通过逐渐降低缓冲液中尿素的浓度在扩张床中进行重折叠过程，然后以填充床模式洗脱回收重折叠的rhIFN-γ蛋白。结果表明，该方法能够高效地重折叠变性的rhIFN-γ蛋白，产量较高。在适当的实验条件下，rhIFN-γ的蛋白产量和比活性分别高达52.7%和8.18×10⁶ IU/mg。
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