Multiplex analysis of enzyme kinetics and inhibition by droplet microfluidics using picoinjectors
2012 (English)In: Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012, 2012, 172-174 p.Conference paper (Refereed)
We present a novel microdroplet-based device for extensive characterization of the reaction kinetics of enzymeinhibitor systems in a single experiment, for the first time utilizing droplet picoinjectors for bioanalysis. This device enables the scanning of multiple inhibitors, inhibitor concentrations and substrate conditions in a single, highly time resolved experiment yielding the Michaelis constant (Km), the turnover number (Kcat) the mode of inhibition and the inhibitor enzyme binding constants (Ki, Ki). Using this device we determine Km and Kcat for β-galactosidase and the fluorogenic substrate Resorufin β-D-galactopyranoside (RBG) to 252 μM and 477 s-1, respectively. Furthermore, we examine the inhibitory effects of Phenylethyl β-D-thiogalactopyranoside (PETG) on this system.
Place, publisher, year, edition, pages
2012. 172-174 p.
Droplet microfluidics, Enzyme inhibitors, Enzyme kinetics, High throughput, Drops, Experiments, Kinetics, Microfluidics, Reaction kinetics, Fluorogenic substrate, Michaelis constants, Multiplex analysis, Substrate conditions, Time resolved experiments, Enzyme inhibition
IdentifiersURN: urn:nbn:se:kth:diva-167650ScopusID: 2-s2.0-84901759186ISBN: 9780979806452OAI: oai:DiVA.org:kth-167650DiVA: diva2:816859
16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012, 28 October-1 November 2012, Okinawa, Japan
QC 201506042015-06-042015-05-222015-06-04Bibliographically approved