Fast and accurate enzyme activity measurements using a chip-based microfluidic calorimeter

Morten van Schie, K. Honarmand Ebrahimi, Wilfred R. Hagen, Peter-Leon Hagedoorn

Research output: Contribution to journalArticleScientificpeer-review

4 Citations (Scopus)
17 Downloads (Pure)

Abstract

Recent developments in microfluidic and nanofluidic technologies have resulted in development of new chip-based microfluidic calorimeters with potential use in different fields. One application would be the accurate high-throughput measurement of enzyme activity. Calorimetry is a generic way to measure activity of enzymes, but unlike conventional calorimeters, chip-based calorimeters can be easily automated and implemented in high-throughput screening platforms. However, application of chip-based microfluidic calorimeters to measure enzyme activity has been limited due to problems associated with miniaturization such as incomplete mixing and a decrease in volumetric heat generated. To address these problems we introduced a calibration method and devised a convenient protocol for using a chip-based microfluidic calorimeter. Using the new calibration method, the progress curve of alkaline phosphatase, which has product inhibition for phosphate, measured by the calorimeter was the same as that recorded by UV-visible spectroscopy. Our results may enable use of current chip-based microfluidic calorimeters in a simple manner as a tool for high-throughput screening of enzyme activity with potential applications in drug discovery and enzyme engineering.
Original languageEnglish
Pages (from-to)57-63
Number of pages7
JournalAnalytical Biochemistry
Volume544
DOIs
Publication statusPublished - Mar 2017

Keywords

  • Chip-based calorimetry
  • Enzyme calorimetry
  • Chemical calibration method
  • Alkaline phosphatase
  • Phosphate ionization

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