PROTEIN REFOLDING BY SIZE EXCLUSION CHROMATOGRAPHY

LU Diannan; LIU Zheng

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PROTEIN REFOLDING BY SIZE EXCLUSION CHROMATOGRAPHY

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- PROTEIN REFOLDING BY SIZE EXCLUSION CHROMATOGRAPHY
- CIESC Journal Issue 10, (2002)
- 作者机构：
  
  清华大学化学工程系
- 作者简介：
- 基金信息：
- DOI：
  CLC：
- Online First：25 October 2002，
  
  Published：25 October 2002
- 稿件说明：
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LU Diannan, LIU Zheng. PROTEIN REFOLDING BY SIZE EXCLUSION CHROMATOGRAPHY[J]. CIESC Journal, 2002, (10).
DOI：

LU Diannan, LIU Zheng. PROTEIN REFOLDING BY SIZE EXCLUSION CHROMATOGRAPHY[J]. CIESC Journal, 2002, (10). DOI：

摘要

以溶菌酶为模拟体系对体积排阻色谱法进行蛋白质折叠过程实验研究 .圆二色性光谱法分析结果证实了复性溶菌酶与天然溶菌酶的二级结构一致性 ;复性溶菌酶与天然溶菌酶色谱保留体积的差异揭示出折叠过程中无活性蛋白质聚集体的存在及其向复性蛋白质转化的机制 ;不同初始浓度的复性实验证实了蛋白质聚集体的存在及其与变性蛋白质初始浓度的关系 ;采用短色谱柱的折叠分离实验结果表明蛋白质折叠是一个快速过程 ;不同尿素浓度下的折叠分离实验结果表明尿素在SEC法中具有非常重要的作用 .与稀释复性法的对比实验表明 :体积排阻色谱法具有稀释倍数小、复性产品活性收率高、复性蛋白质浓度高等优点 .

Abstract

Investigations on protein refolding by size-exclusion chromatography (SEC) was carried out by using lysozyme as a model system.The comparison of elution volume of the native lysozyme and the refolded lysozyme demonstrated the variation of protein diameters during its refolding.The high yield of lysozyme activity obtained at high starting concentration illustrates the workability of this method for high concentration protein refolding.The effects of the sample loading and elution flowrate on the recovery of lysozyme were investigated

separately.The circular dichroism analysis demonstrated the structural identity of the refolded lysozyme and the native lysozyme.Short columns were used for the investigation of the refolding kinetic

and the impact of separation on refolding.The experimental results described above showed the advantages of this new method over the conventional step-wise dilution procedure in terms of high sample concentration and the in situ polishing and refolding.

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