Capillary Electrophoresis in Enzyme Assays and Inhibitor Screening

Capillary electrophoresis (CE) has emerged as a versatile and high‐resolution analytical technique for enzyme kinetics and inhibitor screening. By harnessing narrow‐bore capillaries under high‐voltage fields, CE separates charged molecular species in seconds to minutes, consuming only nanolitre volumes of sample. The intrinsic speed, resolution and minimised reagent consumption of CE make it particularly suited for monitoring enzymatic transformations in real time. Integration of in‐capillary reaction formats—either by immobilising enzyme molecules onto the capillary wall or by mixing reactants through controlled electromigration—has yielded robust enzyme microreactors that streamline assays and enhance throughput.

Such microreactor formats facilitate the direct quantification of reaction products and unreacted substrates, enabling precise determination of kinetic constants and inhibitor potency with exceptional reproducibility. The combination of CE with diverse detection modes, including UV absorption, fluorescence and electrochemiluminescence, further broadens its applicability to a wide spectrum of enzymes and inhibitors. These developments have global significance for drug discovery pipelines, environmental monitoring of enzyme activity and clinical diagnostics, where rapid and reliable assessment of enzyme–inhibitor interactions is essential.

Capillary platforms have also been instrumental in screening libraries of small‐molecule inhibitors, where rapid separation of enzyme–inhibitor complexes or reaction products can be achieved in a single run. The high sensitivity and automation compatibility of CE permit miniaturised screening workflows, reducing reagent costs and accelerating hit identification in early drug discovery.

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Capillary Electrophoresis in Enzyme Assays and Inhibitor Screening publication trend

The graph below shows the total number of articles in capillary electrophoresis in enzyme assays and inhibitor screening across all publications each year (not limited to Nature Index journals).

Technical terms

Capillary electrophoresis (CE): An analytical separation method using narrow capillaries and high voltage to resolve ionic species according to their charge-to-size ratio.

Immobilised enzyme microreactor (IMER): A format in which enzymes are fixed inside a CE capillary to catalyse reactions inline during electrophoretic separation.

Transverse Diffusion of Laminar Flow Profiles (TDLFP): A diffusion-based mixing technique within CE capillaries that enables homogeneous reactant interaction without surface modification.

Electrochemiluminescence detection (ECL): A detection mode where electrochemical excitation induces luminescence, providing high sensitivity in CE analyses.

References

1. Screening of Acetylcholinesterase Inhibitors by Capillary Electrophoresis with Oriented-Immobilized Enzyme Microreactors Based on Gold Nanoparticles. Molecules (2023).
2. Atypical applications of transverse diffusion of laminar flow profiles methodology for in‐capillary reactions in capillary electrophoresis. Journal of Separation Science (2024).
3. Application of capillary electrophoresis coupling with electrochemiluminescence detection to estimate activity of butyrylcholinesterase. Journal of Applied Pharmaceutical Science (2018).