https://scholars.lib.ntu.edu.tw/handle/123456789/61154
標題: | Step change of mobile phase flow rates to enhance protein folding in size exclusion chromatography | 作者: | STEVEN SHENG-SHIH WANG Chang, Che-Kuei HWAI-SHEN LIU |
關鍵字: | Step change of mobile phase flow rates;Chaperon solvent plug;Size exclusion chromatography;Protein refolding;Lysozyme | 公開日期: | 2006 | 出版社: | Taipei:National Taiwan University Dept Chem Engn | 起(迄)頁: | - | 來源出版物: | Biochemical Engineering Journal 29(2006), 2–11 | 摘要: | Minimal aggregation formation between injector and column inlet and enough residence time inside the column were identified to be the key for proper proteins refolding by size exclusion chromatography (SEC). Therefore, a step change of mobile phase flow rate strategy was developed. That is, during the injected denatured protein (sample) traveling from injector to column inlet, a higher rate of mobile phase should be applied to reduce aggregation. Then, a lower flow rate should be used to allow enough time for protein to refold inside the column. The refolding of denatured lysozyme in SEC by this method was shown to reduce the aggregation and thus resulting in good refolding yields, though not completely. Combining this method together with our previously proposed chaperon solvent plug strategy could obtain complete recovery of denatured protein, both mass and activity recoveries. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/2006111501254434 | 其他識別: | 246246/2006111501254434 | DOI: | 10.1016/j.bej.2005.02.036 |
顯示於: | 化學工程學系 |
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