Nilsson Mikael, Harang Valerie, Bergström Maria, Ohlson Sten, Isaksson Roland, Johansson Gunnar
Department of Chemistry and Biomedical Sciences, University of Kalmar, Kalmar, Sweden.
Electrophoresis. 2004 Jun;25(12):1829-36. doi: 10.1002/elps.200405918.
A simple method to calculate dissociation constants for protein-ligand interactions by partial-filling capillary electrophoresis is demonstrated. The method uses raw migration time data for the ligand and needs only additional information about capillary inner radius and the absolute amount of protein loaded. A theoretical study supported by experimental data also demonstrates that the retention of analyte in affinity capillary electrophoresis (ACE) using the partial-filling technique depends linearly on the absolute amount of selector added but is independent of both selector zone length and selector mobility. Factors such as field strength and electroosmotic flow are also cancelled out if they are kept constant. The theory is confirmed and the usefulness of the method is demonstrated by enantioseparations using alpha-acid glycoprotein (AGP) and cellulase (Cel 7A) as chiral selectors.
展示了一种通过部分填充毛细管电泳计算蛋白质-配体相互作用解离常数的简单方法。该方法使用配体的原始迁移时间数据，仅需要关于毛细管内径和加载蛋白质的绝对量的额外信息。一项得到实验数据支持的理论研究还表明，使用部分填充技术的亲和毛细管电泳（ACE）中分析物的保留率与添加的选择剂的绝对量呈线性关系，但与选择剂区带长度和选择剂迁移率均无关。如果场强和电渗流等因素保持恒定，它们也会相互抵消。使用α-酸性糖蛋白（AGP）和纤维素酶（Cel 7A）作为手性选择剂进行对映体分离，证实了该理论并证明了该方法的实用性。
