Capillary Electrophoresis-Based Enzyme Assay for Nicotinamide N-Methyltransferase

Automated High-Throughput System Combining Small-Scale Synthesis with Bioassays and Reaction Screening

“…110 These assays, despite being widely used and even commercially available, have several disadvantages, including the use of radioactive material or labels and the need for multiple preparation steps, which makes them slow and labor-intensive. 82 Recently, methods have been developed around label-free fluorescence assays with a significantly limited substrate scope (only quinolines yielding florescent quinolinium ions upon N -methylation), 82 capillary electrophoresis, 111 and LC (coupled with either UV detection or MS). 83–85,112 Although successful and versatile, the introduction of separation techniques dramatically reduces the throughput of these assays, with chromatographic runs taking 2 min in the fastest cases (using hydrophilic interaction liquid chromatography [HILIC]) 84,85 and up to 25 min in the slowest (using C18 reverse-phase columns).…”

Application of Capillary Electrophoresis with Laser-Induced Fluorescence to Immunoassays and Enzyme Assays

“…The extensive studies of enzymes as current drug targets have encouraged researchers to seek effective methods to characterize enzyme activities and understand their roles in human diseases. Such activities include enzyme kinetics, enzyme substrate identification, enzyme inhibitor screening, and enzyme-mediated metabolic pathways, and these aspects of enzyme-related analysis have been largely studied by CE methods, for example, those summarized in [96,97,98,99]. A typical CE-based enzyme assay starts with small amounts of the substrate, enzyme, and/or inhibitors.…”