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Author: Richter, Andreas × Type: Article ×

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    Hydrogel microvalves as control elements for parallelized enzymatic cascade reactions in microfluidics
    (Basel : MDPI, 2020) Obst, Franziska; Beck, Anthony; Bishayee, Chayan; Mehner, Philipp J.; Richter, Andreas; Voit, Brigitte; Appelhans, Dietmar
    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. © 2020 by the authors.
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    Hydrogel patterns in microfluidic devices by do-it-yourself UV-photolithography suitable for very large-scale integration
    (Basel : MDPI, 2020) Beck, Anthony; Obst, Franziska; Busek, Mathias; Grünzner, Stefan; Mehner, Philipp J.; Paschew, Georgi; Appelhans, Dietmar; Voit, Brigitte; Richter, Andreas
    The interest in large-scale integrated (LSI) microfluidic systems that perform highthroughput biological and chemical laboratory investigations on a single chip is steadily growing. Such highly integrated Labs-on-a-Chip (LoC) provide fast analysis, high functionality, outstanding reproducibility at low cost per sample, and small demand of reagents. One LoC platform technology capable of LSI relies on specific intrinsically active polymers, the so-called stimuli-responsive hydrogels. Analogous to microelectronics, the active components of the chips can be realized by photolithographic micro-patterning of functional layers. The miniaturization potential and the integration degree of the microfluidic circuits depend on the capability of the photolithographic process to pattern hydrogel layers with high resolution, and they typically require expensive cleanroom equipment. Here, we propose, compare, and discuss a cost-efficient do-it-yourself (DIY) photolithographic set-up suitable to micro-pattern hydrogel-layers with a resolution as needed for very large-scale integrated (VLSI) microfluidics. The achievable structure dimensions are in the lower micrometer scale, down to a feature size of 20 µm with aspect ratios of 1:5 and maximum integration densities of 20,000 hydrogel patterns per cm. Furthermore, we demonstrate the effects of miniaturization on the efficiency of a hydrogel-based microreactor system by increasing the surface area to volume (SA:V) ratio of integrated bioactive hydrogels. We then determine and discuss a correlation between ultraviolet (UV) exposure time, cross-linking density of polymers, and the degree of immobilization of bioactive components. © 2020 by the authors.
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    Novel Application of Polymer Networks Carrying Tertiary Amines as a Catalyst Inside Microflow Reactors Used for Knoevenagel Reactions
    (Weinheim : Wiley-VCH Verl., 2020) Berg, Patrik; Obst, Franziska; Simon, David; Richter, Andreas; Appelhans, Dietmar; Kuckling, Dirk
    A novel application is described for utilizing hydrogel dots as organocatalyst carriers inside microfluidic reactors. Tertiary amines were covalently immobilized in the hydrogel dots. Due to the diffusion of reactants within the swollen hydrogel dots, the accessible amount of catalysts inside a microfluidic reactor chamber can be increased compared to the accessible amount of surface-bound catalysts. To perform fast Knoevenagel reactions, important flow parameters had to be validated to optimize the reactor performance while keeping the dimensions of the reactor chamber constant; e.g. the height of the hydrogel dots had to be adjusted to the invariable dimensions of the reactor chamber, or an adjustment of organocatalysts in the hydrogel dots had to be validated to achieve the highest conversion rate during a certain residence time. To characterize the conversion, nuclear magnetic resonance (NMR) and UV/Vis-spectroscopy were utilized as an offline and online method, respectively. With suitable hydrogel dots, the influence of different flow parameters (e.g., operating flow rate and reactant concentration) on the selected model reactions in the microfluidic reactor was investigated. Finally, a variety of reactants were screened with the optimized flow parameters. With these results, the turnover frequency was determined for the Knoevenagel reactions in a microfluidic reactor, and the results were compared with published data that were determined by other synthetic approaches. © 2020 The Authors published by Wiley-VCH GmbH