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首页> 外文会议>The Third China-Korea conference on separation science and technology >Primary Purification of Co-expressed Soluble and Insoluble Alpha-interferon 2b from Recombinant E. coli Homogenate
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Primary Purification of Co-expressed Soluble and Insoluble Alpha-interferon 2b from Recombinant E. coli Homogenate

机译：从重组大肠杆菌匀浆中共表达可溶性和不溶性α-干扰素2b的初步纯化

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摘要

With the development of cloning technology, alpha-interferon 2b (IFN 2b) was produced both in soluble and insoluble forms in recombinant E. coli. The dissolution and refolding of the expressed IFN 2b in inclusion body form were carried out using guanidinium salt. The result showed that 7M guanidinium solution and pH 3.0 were preferred to dissolve the expressed protein, then diluting the resultant solution 15 times at pH 6.0 would lead to correct protein refolding efficiently. The cation exchange column was employed to purify the refolded and soluble IFN 2b respectively. With soluble IFN sample, about 92.1% IFN 2b recovery and 82.2% specific purity were obtained in the elute. However, with refolded IFN sample, only 72.7% IFN 2b recovery and 30.1% specific purity was achieved after cation exchange chromatography. This work laid a base for further purification to meet pharmaceutical product standard.
机译：随着克隆技术的发展，重组大肠杆菌中以可溶和不溶形式生产了α-干扰素2b（IFN 2b）。使用胍盐进行包涵体形式的表达IFN 2b的溶解和重折叠。结果表明，优选使用7M胍溶液和pH 3.0溶解表达的蛋白质，然后将所得溶液在pH 6.0稀释15倍，可以有效地正确折叠蛋白质。阳离子交换柱分别用于纯化重折叠的和可溶的IFN 2b。对于可溶性IFN样品，洗脱液中回收率约为92.1％，比纯度为82.2％。但是，对于重新折叠的IFN样品，阳离子交换色谱分析后仅获得72.7％的IFN 2b回收率和30.1％的比纯度。这项工作为进一步纯化达到药品标准奠定了基础。

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