Hearn M T, Hodder A N, Aguilar M I
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
J Chromatogr. 1988 Dec 23;458:45-56. doi: 10.1016/s0021-9673(00)90552-1.
This investigation deals with protein retention behaviour in high-performance anion-exchange chromatography in terms of the average distance of approach between the protein solute and the positively charged anion-exchange stationary-phase surface. The theoretical treatment is based on a modified Debye-Hückel theory for spherical impenetrable ions, where the electrostatic potential energy has been related to the chromatographic capacity factor, k'. Results are presented for three globular proteins, eluted isocratically from a Mono-Q strong anion-exchange resin with sodium chloride as the displacer salt by a mobile phase with pH in the range 5.50-9.60. Analysis of experimental retention data indicates that topographically predefined, charged regions on the protein surface, called ionotopes, control the orientation and approach distance of the protein solute.
本研究从蛋白质溶质与带正电的阴离子交换固定相表面之间的平均接近距离方面，探讨了高效阴离子交换色谱中的蛋白质保留行为。理论处理基于修正的德拜-休克尔理论，用于球形不可穿透离子，其中静电势能与色谱容量因子k'相关。给出了三种球状蛋白质的结果，这些蛋白质在pH值为5.50 - 9.60的流动相中，以氯化钠作为置换盐，从Mono-Q强阴离子交换树脂上进行等度洗脱。对实验保留数据的分析表明，蛋白质表面在拓扑结构上预先确定的带电荷区域，称为离子位，控制着蛋白质溶质的取向和接近距离。
