用于蛋白同时复性及纯化的制备型装置中流动相用量的优化研究

 在生物大分子的高效液相色谱分离中 ,由计量置换保留模型可得出生物大分子在色谱柱上的保留行为取决于流动相中置换剂的浓度的结论。据此提出了用于蛋白同时复性及纯化的制备型装置 (USRPP)中最小流动相用量的估算公式 ,并进一步得出在保持最小流动相用量不变的条件下 ,改变流动相流速和线性梯度时间几乎不影响制备型USRPP分离蛋白的分离度和复性效率的结论。该结论与实验结果一致。

Study on Optimization of the Required Amount of Mobile Phase for Preparative Unit of Simultaneous Renaturation and Purification of Proteins

Based on the stoichiometric displacement model for retention(SDM R) in high performance liquid chromatography(HPLC), the biopolymer retentions are mainly dominated by displacer's concentration in mobile phase According to this conclusion, an equation is presented for evaluating the minimum amount of mobile phase used for the preparative purpose with the unit of simultaneous renaturation and purification of proteins (USRPP) by linear gradient elution Furthermore, the resolution and refolding efficiency of lysozyme were found to be independent of the flow rate and gradient time when the amount of mobile phase was kept constant The conclusions were tested by experiments and a good coincidence was obtained