Adaptation of a microfluidic qPCR system for enzyme kinetics studies

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Abstract

Microfluidic platforms offer a drastic increase in throughput while minimizing sample usage and hands-on time, which makes them important tools for large-scale biological studies. A range of such systems have been developed for enzyme activity studies, although their complexity largely hinders their application by a wider scientific community. Here we present adaptation of an easy-to-use commercial microfluidic qPCR system for performing enzyme kinetics studies. We demonstrate functionality of the Fluidigm Biomark HD system (the Fluidigm system) by determining kinetic properties of three oxidases in a resorufin-based fluorescent assay. The results obtained in the microfluidic system proved reproducible and comparable to the ones obtained in a standard microplate-based assay. With a wide range of easy-to-use, off-the-shelf components, the microfluidic system presents itself as a simple and customizable platform for high-throughput enzyme activity studies. Graphical abstract

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europepmc
last seen: 2026-05-19T01:45:01.086888+00:00
unpaywall
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License: CC-BY-4.0