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.

中文翻译：

---

通过区域洗脱亲和色谱法分析溶质-蛋白质相互作用和溶质-溶质竞争

许多生物过程涉及溶质-蛋白质相互作用和溶质-溶质竞争蛋白质结合。为检查这些相互作用而开发的一种方法是区域洗脱亲和层析。本综述讨论了区域洗脱亲和色谱的理论和原理及其一般应用。所检查的应用示例包括使用该方法来估计溶质-蛋白质结合的相对程度、检查溶质-溶质竞争和蛋白质的置换，以及测量这些相互作用的强度。它还展示了区域洗脱亲和色谱如何用于溶剂和温度研究以及表征蛋白质上溶质的结合位点。此外，还讨论了区域洗脱亲和色谱的几种替代应用，包括分析溶质与可溶性结合剂的结合以及变构效应的研究。最近考虑的其他应用包括联合使用免疫提取和区域洗脱进行药物-蛋白质结合研究，以及基于固定受体或小靶标的结合研究。