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Mapping of Enzyme Kinetics on a Microfluidic Device.
Hoon Suk Rho1, Alexander Thomas Hanke2, Marcel Ottens2
1Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, Enschede, The Netherlands.
Plos One
|April 16, 2016
Summary
A novel microfluidic mapper enables 36 parallel biochemical reactions in nanoliter volumes. This tool rapidly analyzes enzyme kinetics and optimizes reactions by generating dual concentration gradients for multiple reagent interactions.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Chemical Engineering
Background:
- Microfluidic devices offer miniaturized platforms for chemical and biochemical analyses.
- High-throughput screening of reactions often requires precise control over reagent concentrations.
- Evaluating complex reaction kinetics necessitates efficient methods for exploring multiple conditions simultaneously.
Purpose of the Study:
- To demonstrate a microfluidic platform, termed the "microfluidic mapper," for performing numerous parallel biochemical reactions.
- To showcase the system's capability in generating stepwise dual concentration gradients for two reagents.
- To establish a rapid analytical tool for optimizing chemical and biochemical reaction kinetics.
Main Methods:
- Development of a microfluidic device capable of executing 36 parallel reactions.
- Generation of stepwise concentration gradients for two distinct reagents within each reaction.
- Utilizing a nanoliter-scale reaction volume (36 nl) for each parallel experiment.
- Real-time monitoring of enzymatic reactions, exemplified by horseradish peroxidase (HRP) with Amplex Red (AR) and hydrogen peroxide (H2O2).
Main Results:
- Successful execution of 36 parallel biochemical reactions with varying reagent concentrations.
- Obtained a 3D enzyme reaction plot for HRP, AR, and H2O2 across defined concentration ranges (AR: 11.7–100.0 μM, H2O2: 11.1–66.7 μM).
- Demonstrated the advantages of parallelization and real-time monitoring for rapid gradient generation.
Conclusions:
- The microfluidic mapper serves as a powerful and fast analytical tool for evaluating chemical and biochemical reactions, particularly those involving multiple interacting reagents.
- The system's ability to generate dual concentration gradients in parallel nanoliter-scale reactions facilitates efficient kinetic studies.
- Potential applications include revealing binding kinetics and optimizing reaction conditions in analytical chemistry and related fields.
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