Refolding of protein inclusion bodies directly from E-coli homogenate using expanded bed adsorption chromatography
Refolding of protein inclusion bodies directly from E-coli homogenate using expanded bed adsorption chromatography
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DOI：
10.1023/a:1016305603569
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发表时间：
2001-01-01
期刊：
影响因子：
--
通讯作者：
Lee, EK
中科院分区：
文献类型：
--
作者：
Cho, TH;Ahn, SJ;Lee, EK
为了避免传统溶液相复性过程中固有的聚集问题，我们提出了一种与膨胀床吸附色谱相结合的固相复性方法。模型蛋白是重组人生长激素和谷胱甘肽S-转移酶片段的融合蛋白。结果表明，细胞匀浆中的包涵体蛋白可直接复性，复性率较高。为了验证该方法的适用性，我们成功地测试了三种类型的起始材料，即，重组人生长激素（rhGH）单体、含有融合蛋白的包涵体和E. coli细胞匀浆。这种直接复性方法可以减少所需的复性步骤的数量，并允许在更高的浓度下复性，约2 mg融合蛋白/ml树脂。
To avoid the intrinsic problem of aggregation associated with the traditional solution-phase refolding process, we proposed a solid-phase refolding method integrated with the expanded bed adsorption chromatography. The model protein was a fusion protein of recombinant human growth hormone and a glutathione S-transferase fragment. It was demonstrated that the inclusion body proteins in the cell homogenate could be directly refolded with higher yield. To verify the applicability of this method, we have tested with success three types of the starting materials, i.e., rhGH monomer, inclusion bodies containing the fusion protein, and the E. coli cell homogenate. This direct refolding process could reduce the number of the renaturation steps required and allow the refolding at a higher concentration, approximately 2 mg fusion protein per ml resin.