The study of drug–protein interactions can reveal the corresponding binding mechanisms, providing valuable information for the early phase drug development and development of new drugs. This article reviews the methods used for obtaining the binding parameters of drug–protein systems. The methods include equilibrium dialysis, high-performance affinity chromatography, capillary electrophoresis, spectroscopy, calorimetry, competition and displacement, mass spectrometry, fluorescence resonance energy transfer, and thermal stability shift analysis. Relevant parameters include the association constant, number of binding sites, thermodynamic properties, binding force types, binding site types, binding distances, changes in protein conformation, and changes in protein stability. In addition, the review also summarizes the principles, advantages, and limitations of each method in detail. The comparison of parameter information can not only guide method selection but also provide valuable reference information for in-depth exploration of drug–protein interaction mechanisms.

药物-蛋白质相互作用的研究可以揭示相应的结合机制，为早期药物开发和新药开发提供有价值的信息。本文回顾了用于获得药物-蛋白质系统结合参数的方法。这些方法包括平衡透析、高效亲和色谱、毛细管电泳、光谱学、量热法、竞争和置换、质谱法、荧光共振能量转移和热稳定性偏移分析。相关参数包括缔合常数、结合位点数量、热力学性质、结合力类型、结合位点类型、结合距离、蛋白质构象变化和蛋白质稳定性变化。此外，综述还总结了原则、优势、以及每种方法的局限性。参数信息的比对不仅可以指导方法选择，还可以为深入探索药物-蛋白质相互作用机制提供有价值的参考信息。