Department of Chemistry, University of Nebraska, Lincoln, NE, USA.
Department of Optometry and Vision Science, University of Alabama at Birmingham, Birmingham, AL, USA.
Methods. 2018 Aug 15;146:3-11. doi: 10.1016/j.ymeth.2018.01.020. Epub 2018 Feb 2.
Many biological processes involve solute-protein interactions and solute-solute competition for protein binding. One method that has been developed to examine these interactions is zonal elution affinity chromatography. This review discusses the theory and principles of zonal elution affinity chromatography, along with its general applications. Examples of applications that are examined include the use of this method to estimate the relative extent of solute-protein binding, to examine solute-solute competition and displacement from proteins, and to measure the strength of these interactions. It is also shown how zonal elution affinity chromatography can be used in solvent and temperature studies and to characterize the binding sites for solutes on proteins. In addition, several alternative applications of zonal elution affinity chromatography are discussed, which include the analysis of binding by a solute with a soluble binding agent and studies of allosteric effects. Other recent applications that are considered are the combined use of immunoextraction and zonal elution for drug-protein binding studies, and binding studies that are based on immobilized receptors or small targets.
许多生物过程涉及溶质-蛋白质相互作用和溶质-溶质对蛋白质结合的竞争。已经开发出一种方法来检查这些相互作用，即区带洗脱亲和层析。本文讨论了区带洗脱亲和层析的理论和原理，以及它的一般应用。检查的应用实例包括使用该方法估计溶质-蛋白质结合的相对程度、检查溶质-溶质竞争和从蛋白质中置换，以及测量这些相互作用的强度。还展示了如何在溶剂和温度研究中使用区带洗脱亲和层析，并描述溶质在蛋白质上的结合位点。此外，还讨论了区带洗脱亲和层析的其他几种替代应用，包括与可溶性结合剂结合的溶质的分析以及变构效应的研究。考虑的其他最近的应用包括免疫提取与区带洗脱联合用于药物-蛋白质结合研究，以及基于固定化受体或小靶标的结合研究。
