Available online 24 July 2025, 101407

Affinity chromatography can be used in many ways to study drug-target interactions.

Zonal elution and frontal analysis methods are often used for this purpose.

Various affinity chromatographic methods for kinetic studies are also available.

Secondary capture agents or hybrid affinity methods can also be used.

The applications, advantages and limitations for each of these methods are discussed.

The characterization of drug-target interactions is a key component of drug discovery, testing, and development. Affinity chromatography is one approach that can be used for this type of analysis. For instance, this may be done by using an immobilized target as a stationary phase and a drug as the applied solute. This review will discuss the various ways in which affinity chromatographic methods have been used to examine drug-target interactions, with an emphasis on high-performance methods. The general principles of this approach and factors to consider in its use for drug-target interaction analysis will first be examined. Methods based on zonal elution or frontal analysis for binding and competition studies will then be discussed. Various techniques for kinetic studies will next be considered, along with approaches that employ secondary binding agents and hybrid techniques. In each case, the general principles and theory of an approach will be given along with examples of its use in drug-target interaction studies. Advantages or limitations of each approach will be provided as well. This information should make it possible in the future to extend these techniques to other drug-target systems of interest in biomedical research and drug testing or development.

Affinity chromatography

High-performance affinity chromatography

Drug-target interactions

Drug-Protein binding

Kinetic analysis

Biointeraction analysis

© 2025 The Authors. Published by Elsevier B.V. on behalf of Xi’an Jiaotong University.