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首页> 外文期刊>Biotechnology Progress >Mechanism of pH-Sensitive Polymer-Assisted Protein Refolding and Its Application in TGF-β1 and KGF-2
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Mechanism of pH-Sensitive Polymer-Assisted Protein Refolding and Its Application in TGF-β1 and KGF-2

机译：pH敏感的聚合物辅助蛋白复性机理及其在TGF-β1和KGF-2中的应用

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摘要

Refolding of proteins at high concentrations often results in non-productive aggregation. This study, through a unique combination of spectroscopic and chromatographic analyzes, provides biomolecular evidence to demonstrate the ability of Eudragit S-100, a pH-respon-sive polymer, to enhance refolding of denatured-reduced lysozyme at high concentrations. The addition of Eudragit in the refolding buffer significantly increases lysozyme refolding yield to 75%, when dilution refolding was conducted at 1 mg/mL lysozyme. This study shows evidence of an electrostatic interaction between oppositely charged lysozyme and the Eudragit polymer during refolding. This ionic complexing of Eudragit and lysozyme appears to shield exposed hydrophobic residues of the lysozyme refolding intermediates, thus minimizing hydrophobic-driven aggregation of the molecules. Importantly, results from this study show that the Eudragit-lysozyme bioconjugation does not compromise refolded protein structure, and that the polymer can be readily dissociated from the protein by ion exchange chromatography. The strategy was also applied to refolding of TGF-β1 and KGF-2.
机译：高浓度蛋白质的折叠通常会导致非生产性聚集。这项研究通过光谱分析和色谱分析的独特结合，提供了生物分子证据来证明具有pH响应性的Eudragit S-100在高浓度下增强变性还原溶菌酶重折叠的能力。当以1 mg / mL溶菌酶进行稀释重折叠时，在重折叠缓冲液中添加Eudragit可以显着提高溶菌酶的重折叠产率至75％。这项研究表明，在重新折叠期间，带相反电荷的溶菌酶和Eudragit聚合物之间存在静电相互作用的证据。 Eudragit和溶菌酶的这种离子络合似乎屏蔽了溶菌酶折叠中间体的暴露的疏水残基，从而使分子的疏水驱动聚集最小化。重要的是，这项研究的结果表明，Eudragit-溶菌酶的生物共轭作用不会损害重新折叠的蛋白质结构，并且可以通过离子交换色谱法轻易地将聚合物从蛋白质上解离。该策略还应用于TGF-β1和KGF-2的重折叠。

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