Enzyme kinetics studies play important role in understanding the mechanism of enzyme. The effect of different conditions on the reaction rate is measured. The active compound can inhibit enzyme using three different mechanisms, and enzyme kinetic measurements can elucidate whether it binds to the active site, in its close proximity or to the allosteric site. Different mechanisms will result in different relationship between the compound concertation and observed equilibrium constant. Our microfluidic platform allows to study enzyme kinetics and inhibitor mechanism of action, producing highly precise and reproducible results. Kinetic analysis is achievable due to visualization of product and substrate in ‘real-time’.
Kinetic studies carried out on a microfluidic platform benefit from:
- Ratiometric, simultaneous measurement of substrate and product drives higher precision
- Minimal interference from fluorescent compounds
- Successfully used with different classes of enzymes, broad range of targets including kinases, proteases, epigenetic targets, etc
- The reaction mixture can be sipped and separated at different time points to give a real-time kinetic measurement
Bibliography
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- Yichin Liu, Raphaele Gerber, John Wu, Trace Tsuruda, John D. McCarter, High-throughput assays for sirtuin enzymes: A microfluidic mobility shift assay and a bioluminescence assay, In Analytical Biochemistry, Volume 378, Issue 1, 2008, Pages 53-59, ISSN 0003-2697. Google Scholar