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Effect of ionic capacity on dynamic adsorption behavior of protein in ion-exchange electrochromatography

机译：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

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Four anion-exchangers of different ion-exchange capacities (21 -129 μ mol/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 186 mg/mL in the ionic capacity range, but the effective pore diffusion coefficient of the protein decreased from 10.9 x 10~(-12) to 1.85 x 10~(-12) m2/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 30 mA, the DBC in IEEC packed with ion-exchangers of high ionic capacities (53 and 129 μ mol/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 35 μ mol/mL) increased only slightly (ca. 10%) under the same condition. The results indicated the minor effect of elec-trophoretic 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.
机译：通过将二乙氨基乙基偶联到Sepharose 6FF上制备了四个不同离子交换容量（21 -129μmol / mL）的阴离子交换剂。吸附剂用于研究离子交换电色谱法中动态蛋白质的吸附，其中垂直于流动相流（IEEC）的振荡横向电场。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，但蛋白质的有效孔扩散系数从10.9 x 10〜（-12）降低至1.85 x 10〜 -12）m2 / s具有增加的离子容量，这是由于结合蛋白分子在孔口处的阻碍作用所致。因此，离子交换色谱中蛋白质的动态结合容量（DBC）随着离子容量的增加而降低。通过施加30 mA的电场，IEEC中装有高离子容量（53和129μmol / mL离子交换器）的DBC显着增加（分别超过30％和100％）。这是因为高表面电荷密度导致高电渗流，从而增强了IEEC中的颗粒内质量传递。相比之下，在相同条件下，装有低离子容量（21和35μmol / mL）离子交换剂的IEEC中的DBC仅略有增加（约10％）。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对通过IEEC的蛋白质的高容量纯化是有利的。

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