横向电场体积排阻电色谱中蛋白质保留特性的实验研讨.pdfVIP

L040 横向电场体积排阻电色谱中蛋白质保留特性的实验 研究 * 谭国民 史清洪 孙彦 （天津大学化工学院生物化工系，天津，300072） 摘要：为了强化色谱介质的孔内传质，本文研究了牛血清白蛋白（BSA）和肌红蛋白（Myo）在新设计 的横向电场体积排阻电色谱柱中的保留行为，并考察了缓冲液pH、流速和电流变化周期对保留行为的 影响。实验结果显示尽管Myo 和BSA表观容量因子增量都随电流强度的增加而增加，但电流强度对BSA 的影响更加显著。此外，缓冲液的pH、离子强度的改变将影响蛋白质的zeta电势和/或溶液电导率， 从而改变蛋白质在电色谱中的保留行为。交变电场周期也对蛋白质的保留行为有着显著的影响。最后， 电色谱用以进行BSA和 Myo 混合物分离的尝试。实验表明，外加电场的引入可以增大BSA和Myo 之间 的分离度，并使其得到部分分离。 关键词：体积排阻色谱 电泳迁移 浓差极化 交变电场 制备色谱 EXPERIMENTAL STUDIES OF PROTEINS RETENTION IN SIZE-EXCLUSION ELECTROCHROMATOGRAPHY UNDER TRANSVERSE ELECTRIC FIELD TAN Guomin, SHI Qinghong and SUN Yan (School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China) Abstract A novel three-compartment chromatographic column was designed for electrochromatography of proteins under transverse electric field. The size exclusion chromatographic media was packed in the central compartment partitioned off neighboring electrode compartments by ceramic plate filling with polyacrylamide gel. The retention behavior of bovine serum albumin (BSA) and myoglobin (Myo) in size exclusion electrochromatgraphy (SEEC) was investigated in detail, including the effect of buffer pH, ionic strength and current cycle time. The results showed that the augmented apparent capacity factors for both proteins increased with a elevated current density, in particularly for BSA. Moreover, the zeta potential of protein and conductivity in buffer will change with pH value and ionic strength in buffer, further influence the retention time of protein in electrochromatography. Finally, the attempt on separat