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Multiplex analysis of enzyme kinetics and inhibition by droplet microfluidics using picoinjectors
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.ORCID iD: 0000-0001-5232-0805
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
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, Published paper (Refereed)
Abstract [en]

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.
Keyword [en]
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
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:kth:diva-167650Scopus ID: 2-s2.0-84901759186ISBN: 9780979806452 (print)OAI: oai:DiVA.org:kth-167650DiVA: diva2:816859
Conference
16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012, 28 October-1 November 2012, Okinawa, Japan
Note

QC 20150604

Available from: 2015-06-04 Created: 2015-05-22 Last updated: 2015-06-04Bibliographically approved

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Jönsson, Håkan N.

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