Analysis of solution-phase biomolecular interactions by liquid chromatography: General strategies and recent
David S Hage1, Sadia Sharmeen1, Kyungah Suh1
1Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE 68588-0304, USA.
Journal of Pharmaceutical and Biomedical Analysis
|December 14, 2024
Summary
Liquid chromatography (LC) and high-performance liquid chromatography (HPLC) offer versatile strategies for analyzing biomolecular interactions in solution. These methods enable detailed characterization of binding kinetics and extents for biological processes.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biophysics
Background:
- Biomolecular interactions are crucial for biological system functions.
- Studying these interactions requires methods to analyze binding extent and rate.
- Both homogeneous (solution-phase) and heterogeneous (solution and solid-phase) techniques exist.
Purpose of the Study:
- To review strategies for using LC and HPLC to study biomolecular interactions.
- To explore various LC/HPLC-based approaches for analyzing solution-phase binding.
- To discuss the principles, advantages, limitations, and applications of these methods.
Main Methods:
- Hummel-Dreyer method
- Solution-phase frontal analysis
- Physical entrapment techniques
- Affinity chromatography using stationary phase as capture agent
- Ultrafast affinity extraction
Main Results:
- LC and HPLC can be adapted to study solution-phase biomolecular interactions.
- Multiple strategies leverage chromatographic principles for binding analysis.
- Each method offers unique benefits and drawbacks for specific applications.
Conclusions:
- LC and HPLC provide powerful tools for dissecting biomolecular interactions.
- The reviewed strategies offer flexibility in studying binding kinetics and thermodynamics.
- These chromatographic approaches enhance the understanding of fundamental biological processes.
Keywords:
Biomolecular interactionsEntrapmentFrontal analysisHummel-Dreyer methodNormal-role affinity chromatographyUltrafast affinity extractionMore Related Videos
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