Oscillatory transverse electric field enhances mass transfer and protein capacity in ion-exchange electrochromatography.
Oscillatory transverse electric field enhances mass transfer and protein capacity in ion-exchange electrochromatography.
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DOI：
10.1016/j.chroma.2005.08.089
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发表时间：
2005-12
期刊：
影响因子：
--
通讯作者：
Guo-Min Tan;Qinghong Shi;Y. Sun
中科院分区：
文献类型：
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作者：
Guo-Min Tan;Qinghong Shi;Y. Sun
建立了垂直于压力驱动流动相流动的振荡电场离子交换电色谱（pIEEC）。在pH8.2的Tris-甘氨酸缓冲液中，通过前沿分析研究了电场强度对牛血清白蛋白（BSA）在DEAESepharoseFF填充床上吸附动态结合容量（DBC）的影响。结果表明，pIEEC中10%穿透（Q10）处的DBC随电场强度的增加而线性增加。例如，当填充床高度为15 mm，电位梯度为38 V/cm时，Q_（10）比常规离子交换色谱提高了4倍。因此，横向电场产生了显著的电动质量传输（电渗和电泳），其增强了外部液膜和颗粒内的质量传输，导致蛋白质结合能力增加。由于pIEEC的容量增加，在过载条件下实现了BSA和IgG的部分分离，而无需任何工艺优化。结果表明，20 V/cm的电位梯度足以大大提高pIEEC中的DBC，并且在必要时，可以用低的施加电压实现高电场强度，因为柱的侧距离通常比其高度小一个数量级。使用低电压进行电色谱是pIEEC相对于具有轴向电场的常规电色谱的显著优点。
Ion-exchange electrochromatography with an oscillatory electric field perpendicular to mobile-phase flow driven by pressure (pIEEC) was developed with a column design of rectangle cross-section. The effect of electric field strength on the dynamic binding capacity (DBC) was examined by frontal analysis of bovine serum albumin (BSA) adsorption to the packed beds of DEAE Sepharose FF in Tris–glycine buffer (pH 8.2). It was shown that the DBC at 10% breakthrough (Q10) in the pIEEC increased linearly with increasing the electric field strength. For example, with a packed-bed height of 15mm and electric potential gradient of 38V/cm, Q10increased four times over that in normal ion-exchange chromatography. So, the transverse electric field has created significant electro-kinetic mass transports (electroosmosis and electrophoresis) that intensified exterior liquid-film and intraparticle mass transfers, leading to the increased protein binding capacity. Due to the increased capacity in the pIEEC, partial resolution of BSA and IgG under an overload condition was realized without any process optimization. The results have revealed that an electric potential gradient of 20V/cm was enough to greatly enhance the DBC in the pIEEC, and when necessary, high electric field strength can be realized with a low applied voltage because the side distance of the column is usually an order of magnitude smaller than its height. The use of low voltage to carry out electrochromatography is a significant advantage of the pIEEC over conventional electrochromatography with axial electric field.