Effect of ionic capacity on dynamic adsorption behavior of protein in ion-exchange electrochromatography
Effect of ionic capacity on dynamic adsorption behavior of protein in ion-exchange electrochromatography
复制标题
离子交换电色谱中离子容量对蛋白质动态吸附行为的影响
DOI：
10.1016/j.seppur.2009.04.002
复制
发表时间：
2009-06
影响因子：
8.6
通讯作者：
中科院分区：
文献类型：
--
作者：
将二乙基氨基乙基偶联到Sepharose 6 FF上，制备了四种不同离子交换容量（21-129μmol/mL）的阴离子交换剂。研究了在垂直于流动相流方向的横向振荡电场作用下，该吸附剂对蛋白质的动态吸附行为。在离子容量范围内，牛血清白蛋白（BSA）的静态吸附容量从67 mg/mL增加到186 mg/mL，但由于结合蛋白分子在孔入口处的阻碍作用，BSA的有效孔扩散系数从10.9×10− 12 m2/s降低到1.85×10− 12 m2/s。因此，蛋白质在离子交换层析中的动态结合容量（DBC）随离子容量的增大而减小。在30 mA的电场作用下，高离子容量（53 μ mol/mL和129μmol/mL）离子交换剂填充的IEEC中的DBC分别显著增加30%和100%以上。这是因为高的表面电荷密度导致高的电渗流，增强了IEEC中的粒内传质。相比之下，填充低离子容量（21和35μmol/mL）的离子交换剂的IEEC中的DBC仅略微增加（约2.5 μmol/mL）。10%）在相同条件下。结果表明，电泳迁移率对颗粒内传质的影响较小。因此，使用高离子容量的离子交换剂通过IEEC进行蛋白质的高容量纯化是有益的。
Four anion-exchangers of different ion-exchange capacities (21–129μmol/mL) were prepared by coupling diethylaminoethyl to Sepharose 6FF. The adsorbents were used to study the dynamic protein adsorption in ion-exchange electrochromatography with an oscillatory transverse electric field perpendicular to the mobile-phase flow (IEEC). The static adsorption capacity of bovine serum albumin (BSA) increased from 67 to 186mg/mL in the ionic capacity range, but the effective pore diffusion coefficient of the protein decreased from 10.9×10−12to 1.85×10−12m2/s with increasing the ionic capacity due to the hindrance effect of the bound protein molecules at the pore entrance. So, the dynamic binding capacity (DBC) of protein in ion-exchange chromatography decreased with increasing the ionic capacity. By applying an electric field of 30mA, the DBC in IEEC packed with ion-exchangers of high ionic capacities (53 and 129μmol/mL) increased significantly (over 30% and 100%, respectively). This was because the high surface charge density led to high electroosmotic flow that enhanced intraparticle mass transfer in IEEC. In comparison, the DBC in IEEC packed with ion-exchangers of low ionic capacities (21 and 35μmol/mL) increased only slightly (ca. 10%) under the same condition. The results indicated the minor effect of electrophoretic mobility on the intraparticle mass transfer. Hence, it is beneficial to use ion-exchangers of high ionic capacity for high-capacity purification of proteins by IEEC.
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DOI：
10.1016/j.chroma.2005.08.089
发表时间：
2005-12
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Guo-Min Tan;Qinghong Shi;Y. Sun
通讯作者：
Guo-Min Tan;Qinghong Shi;Y. Sun
影响因子：
2.9
作者：
Guo-Min Tan;Qinghong Shi;Y. Sun
通讯作者：
Guo-Min Tan;Qinghong Shi;Y. Sun
影响因子：
3.7
作者：
S. Rudge;S. Basak;M. Ladisch
通讯作者：
S. Rudge;S. Basak;M. Ladisch
影响因子：
2.9
作者：
K. Cole
通讯作者：
K. Cole
影响因子：
4.1
作者：
Cikalo, MG;Bartle, KD;Myers, P
通讯作者：
Myers, P