通过尺寸排阻色谱法对促肾上腺皮质激素释放因子受体1胞外域进行柱上重折叠。
On-column refolding of corticotropin-releasing factor receptor 1 extracellular domain by size exclusion chromatography.
作者信息
Ersoy Maria, Matic Sandra, Schmidt Stefan R
机构信息
Global Protein Science and Supply, AstraZeneca R&D Södertälje, SE-15185 Södertälje, Sweden.
出版信息
Biomed Chromatogr. 2009 Oct;23(10):1121-8. doi: 10.1002/bmc.1233.
Expressing the extracellular domain of corticotropin releasing factor receptor 1 in Escherichia coli usually results in the formation of inclusion bodies. Here we describe the optimization of refolding by applying size exclusion chromatography with a denaturing guanidine hydrochloride gradient and a refolding buffer containing glycerol. Several chromatographic parameters like gradient length, flow rate, sample concentration and chromatography resin characteristics were evaluated. Recovery yields of refolded protein above 50% using a Superdex 200 column demonstrate the usefulness of this method.
在大肠杆菌中表达促肾上腺皮质激素释放因子受体1的细胞外结构域通常会导致包涵体的形成。在此，我们描述了通过应用含有变性盐酸胍梯度的尺寸排阻色谱法和含有甘油的复性缓冲液来优化复性过程。评估了几个色谱参数，如梯度长度、流速、样品浓度和色谱树脂特性。使用Superdex 200柱时，复性蛋白的回收率超过50%，证明了该方法的有效性。
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