Applications of on‐line weak affinity interactions in free solution capillary electrophoresis
Applications of on‐line weak affinity interactions in free solution capillary electrophoresis
复制标题
DOI：
10.1002/1522-2683(200203)23:6
复制
发表时间：
2002-03
期刊：
影响因子：
2.9
通讯作者：
N. Heegaard;M. Nissen;D. Chen
中科院分区：
文献类型：
--
作者：
N. Heegaard;M. Nissen;D. Chen
在某些情况下，当将与一种或多种分离的分析物进行弱亲和相互作用的配体或添加剂加入电泳缓冲液中时，毛细管电泳提供的令人印象深刻的选择性可以进一步增加。当复合分子的迁移不同于自由分子的迁移且分离条件为非变性时，这种在线亲和毛细管电泳方法是可行的。在这篇综述中，我们专注于应用弱相互作用作为工具，以提高密切相关的分子，例如，药物对映体和使用毛细管电泳定量表征这种相互作用。我们描述了结合等温线的方程，说明如何选择性可以通过改变添加剂的浓度来操纵，并显示如何使用的方法来估计结合常数。在线亲和毛细管电泳方法对于对映体分离和生物样品内容物的功能表征特别有价值，这些生物样品的含量仅为微量。
The impressive selectivity offered by capillary electrophoresis can in some cases be further increased when ligands or additives that engage in weak affinity interactions with one or more of the separated analytes are added to the electrophoresis buffer. This on‐line affinity capillary electrophoresis approach is feasible when the migration of complexed molecules is different from the migration of free molecules and when separation conditions are nondenaturing. In this review, we focus on applying weak interactions as tools to enhance the separation of closely related molecules, e.g., drug enantiomers and on using capillary electrophoresis to characterize such interactions quantitatively. We describe the equations for binding isotherms, illustrate how selectivity can be manipulated by varying the additive concentrations, and show how the methods may be used to estimate binding constants. On‐line affinity capillary electrophoresis methods are especially valuable for enantiomeric separations and for functional characterization of the contents of biological samples that are only available in minute quantities.