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

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

van Schie, M.M.C.H. (TU Delft BT/Biocatalysis)
Honarmand Ebrahimi, K. (University of Oxford)
Hagen, Wilfred R.
Hagedoorn, P.L. (TU Delft BT/Biocatalysis)

2017-03

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.

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

http://resolver.tudelft.nl/uuid:2ab7be08-5469-44cd-bf97-ec8c772c32c2

https://doi.org/10.1016/j.ab.2017.12.028

0003-2697

Analytical Biochemistry, 544, 57-63

Institutional Repository

journal article

© 2017 M.M.C.H. van Schie, K. Honarmand Ebrahimi, Wilfred R. Hagen, P.L. Hagedoorn