title: Mapping of Enzyme Kinetics on a Microfluidic Device.
abstract: A microfluidic platform or "microfluidic mapper" is demonstrated, which in a single experiment performs 36 parallel biochemical reactions with 36 different combinations of two reagents in stepwise concentration gradients. The volume used in each individual reaction was 36 nl. With the microfluidic mapper, we obtained a 3D enzyme reaction plot of horseradish peroxidase (HRP) with Amplex Red (AR) and hydrogen peroxide (H2O2), for concentration ranges of 11.7 μM to 100.0 μM and 11.1 μM to 66.7 μM for AR and H2O2, respectively. This system and methodology could be used as a fast analytical tool to evaluate various chemical and biochemical reactions especially where two or more reagents interact with each other. The generation of dual concentration gradients in the present format has many advantages such as parallelization of reactions in a nanoliter-scale volume and the real-time monitoring of processes leading to quick concentration gradients. The microfluidic mapper could be applied to various problems in analytical chemistry such as revealing of binding kinetics, and optimization of reaction kinetics.
authors.affiliation: Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, Enschede, The Netherlands.
authors.forename: Hoon Suk
authors.initials: HS
authors.lastname: Rho
authors.affiliation: BioProcess Engineering group, Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, Delft, The Netherlands.
authors.forename: Alexander Thomas
authors.initials: AT
authors.lastname: Hanke
authors.affiliation: BioProcess Engineering group, Department of Biotechnology, Faculty of Applied Sciences, Delft University of Technology, Delft, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Ottens
authors.affiliation: Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, Enschede, The Netherlands.
authors.forename: Han
authors.initials: H
authors.lastname: Gardeniers
mesh_terms: D002138:Calibration; D004867:Equipment Design; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D021621:Imaging, Three-Dimensional; D007700:Kinetics; D056656:Lab-On-A-Chip Devices / Q000592:standards; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods / Q000592:standards; D008856:Microscopy, Fluorescence; D059008:Time-Lapse Imaging
chemical_list: D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase