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Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics

Compartmentalized microfluidic devices with immobilized catalysts are a valuable tool for overcoming the incompatibility challenge in (bio) catalytic cascade reactions and high-throughput screening of multiple reaction parameters. To achieve flow control in microfluidics, stimuli-responsive hydrogel...

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Autores principales: Obst, Franziska, Beck, Anthony, Bishayee, Chayan, Mehner, Philipp J., Richter, Andreas, Voit, Brigitte, Appelhans, Dietmar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074747/
https://www.ncbi.nlm.nih.gov/pubmed/32033413
http://dx.doi.org/10.3390/mi11020167
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author Obst, Franziska
Beck, Anthony
Bishayee, Chayan
Mehner, Philipp J.
Richter, Andreas
Voit, Brigitte
Appelhans, Dietmar
author_facet Obst, Franziska
Beck, Anthony
Bishayee, Chayan
Mehner, Philipp J.
Richter, Andreas
Voit, Brigitte
Appelhans, Dietmar
author_sort Obst, Franziska
collection PubMed
description Compartmentalized microfluidic devices with immobilized catalysts are a valuable tool for overcoming the incompatibility challenge in (bio) catalytic cascade reactions and high-throughput screening of multiple reaction parameters. To achieve flow control in microfluidics, stimuli-responsive hydrogel microvalves were previously introduced. However, an application of this valve concept for the control of multistep reactions was not yet shown. To fill this gap, we show the integration of thermoresponsive poly(N-isopropylacrylamide) (PNiPAAm) microvalves (diameter: 500 and 600 µm) into PDMS-on-glass microfluidic devices for the control of parallelized enzyme-catalyzed cascade reactions. As a proof-of-principle, the biocatalysts glucose oxidase (GOx), horseradish peroxidase (HRP) and myoglobin (Myo) were immobilized in photopatterned hydrogel dot arrays (diameter of the dots: 350 µm, amount of enzymes: 0.13–2.3 µg) within three compartments of the device. Switching of the microvalves was achieved within 4 to 6 s and thereby the fluid pathway of the enzyme substrate solution (5 mmol/L) in the device was determined. Consequently, either the enzyme cascade reaction GOx-HRP or GOx-Myo was performed and continuously quantified by ultraviolet-visible (UV-Vis) spectroscopy. The functionality of the microvalves was shown in four hourly switching cycles and visualized by the path-dependent substrate conversion.
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spelling pubmed-70747472020-03-20 Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics Obst, Franziska Beck, Anthony Bishayee, Chayan Mehner, Philipp J. Richter, Andreas Voit, Brigitte Appelhans, Dietmar Micromachines (Basel) Article Compartmentalized microfluidic devices with immobilized catalysts are a valuable tool for overcoming the incompatibility challenge in (bio) catalytic cascade reactions and high-throughput screening of multiple reaction parameters. To achieve flow control in microfluidics, stimuli-responsive hydrogel microvalves were previously introduced. However, an application of this valve concept for the control of multistep reactions was not yet shown. To fill this gap, we show the integration of thermoresponsive poly(N-isopropylacrylamide) (PNiPAAm) microvalves (diameter: 500 and 600 µm) into PDMS-on-glass microfluidic devices for the control of parallelized enzyme-catalyzed cascade reactions. As a proof-of-principle, the biocatalysts glucose oxidase (GOx), horseradish peroxidase (HRP) and myoglobin (Myo) were immobilized in photopatterned hydrogel dot arrays (diameter of the dots: 350 µm, amount of enzymes: 0.13–2.3 µg) within three compartments of the device. Switching of the microvalves was achieved within 4 to 6 s and thereby the fluid pathway of the enzyme substrate solution (5 mmol/L) in the device was determined. Consequently, either the enzyme cascade reaction GOx-HRP or GOx-Myo was performed and continuously quantified by ultraviolet-visible (UV-Vis) spectroscopy. The functionality of the microvalves was shown in four hourly switching cycles and visualized by the path-dependent substrate conversion. MDPI 2020-02-05 /pmc/articles/PMC7074747/ /pubmed/32033413 http://dx.doi.org/10.3390/mi11020167 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Obst, Franziska
Beck, Anthony
Bishayee, Chayan
Mehner, Philipp J.
Richter, Andreas
Voit, Brigitte
Appelhans, Dietmar
Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics
title Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics
title_full Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics
title_fullStr Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics
title_full_unstemmed Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics
title_short Hydrogel Microvalves as Control Elements for Parallelized Enzymatic Cascade Reactions in Microfluidics
title_sort hydrogel microvalves as control elements for parallelized enzymatic cascade reactions in microfluidics
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7074747/
https://www.ncbi.nlm.nih.gov/pubmed/32033413
http://dx.doi.org/10.3390/mi11020167
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