An Immobilized Enzyme Reactor for Spatiotemporal Control over Reaction Products

Grant, Jennifer; Modica, Justin A.; Roll, Juliet; Perkovich, Paul; Mrksich, Milan

doi:10.1002/smll.201800923

Cited by 13 publications

(9 citation statements)

References 57 publications

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“…We cast the PDMS block from 3D printed masters to avoid the use of a cleanroom and to enable rapid design prototyping (Figure S2). 13 Separate solutions of phenylglyoxal and the peptide, each at a concentration of 2 mM, were simultaneously injected into separate inlets and allowed to diffusively mix at the base of a Y-junction, where they continued to react as the solution flowed along a single channel that was 340 mm long, 550 μm wide, and 250 μm tall. In the flow, we included a peptide (0.25 mM) lacking an N-terminal cysteine that is unable to undergo internal rearrangement to form the permanent hydroxy-amide bond.…”

Section: Resultsmentioning

confidence: 99%

Using Microfluidics and Imaging SAMDI-MS To Characterize Reaction Kinetics

et al. 2019

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Microfluidic platforms have enabled the simplification of biochemical assays with a significant reduction in the use of reagents, yet the current methods available for analyzing reaction products can limit applications of these approaches. This paper demonstrates a simple microfluidic device that incorporates a functionalized self-assembled monolayer to measure the rate constant for a chemical reaction. The device mixes the reactants and allows them to selectively immobilize to the monolayer at the base of a microfluidic channel in a time-dependent manner as they flow down the channel. Imaging self-assembled monolayers for matrix-assisted laser desorption/ionization mass spectrometry (iSAMDI-MS) is used to acquire a quantitative image representing the time-resolved progress of the reaction as it flowed through the channel. Knowledge of the surface immobilization chemistry and the fluid front characteristics allows for the determination of the chemical reaction rate constant. This approach widens the applicability of microfluidics for chemical reaction monitoring and establishes a label-free method for studying processes that occur within a dispersive regime.

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Section: Resultsmentioning

confidence: 99%

Using Microfluidics and Imaging SAMDI-MS To Characterize Reaction Kinetics

et al. 2019

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“…The files were converted to stl format and printed in a digital printing mode using a Stratasys Connex 350 3D printer in VeroWhite material (Stratasys Direct) with a glossy finish. The 3D printed masters were prepared for PDMS polymerization as previously described . PDMS prepolymer mixture was mixed in a 1:10 ratio (w/w curing agent to prepolymer), degassed in a vacuum desiccator for 15 min, and poured into the 3D printed master.…”

Section: Materials and Methodsmentioning

confidence: 99%

“…The 3D printed masters were prepared for PDMS polymerization as previously described. 9 PDMS prepolymer mixture was mixed in a 1:10 ratio (w/w curing agent to prepolymer), degassed in a vacuum desiccator for 15 min, and poured into the 3D printed master. The master containing PDMS was degassed in a vacuum desiccator for 15 min and placed in a 43 °C oven overnight.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

High-Throughput Enzyme Kinetics with 3D Microfluidics and Imaging SAMDI Mass Spectrometry

Grant¹,

Goudarzi²,

Mrksich³

2018

Anal. Chem.

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Microfluidic systems are important for performing precise reagent manipulations and reducing material consumption in biological assays. However, optical detection methods limit analyses to fluorescent or UV-active compounds and traditional 2D fluidic designs have limited degrees of freedom. This article describes a microfluidic device that has three inputs and performs 2592 distinct enzyme reactions using only 150 μL of reagent with quantitative characterization. This article also introduces imaging self-assembled monolayers for matrix-assisted laser desorption/ionization mass spectrometry (iSAMDI-MS) to map reaction progress, by immobilization of the product onto the floor of the microfluidic channel, into an image that is used for calculating the Michaelis constant (K m). This approach expands the scope of imaging mass spectrometry, microfluidic detection strategies, and the design of high-throughput reaction systems.

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“…Applications in printing biomimetic organoids have shown success [ 93 ] and adaptation to enzyme immobilization is likely to happen soon. Meanwhile, rapid biotechnology developments are making it easier to modify native enzymes to include beneficial binding domains [ 31 ] that interact or bond to the 3D printed surfaces spontaneously [ 94 ]. This will allow future development of reusable 3D printed devices that can be re-immobilized with enzymes after initial activity loss with a single dip of the 3D structure or create highly targeted constructs by VP techniques with minimal loss of enzymes in the vat phase.…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Advances in 3D Gel Printing for Enzyme Immobilization

Shen

Zhang

Fang

et al. 2022

Gels

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Incorporating enzymes with three-dimensional (3D) printing is an exciting new field of convergence research that holds infinite potential for creating highly customizable components with diverse and efficient biocatalytic properties. Enzymes, nature’s nanoscale protein-based catalysts, perform crucial functions in biological systems and play increasingly important roles in modern chemical processing methods, cascade reactions, and sensor technologies. Immobilizing enzymes on solid carriers facilitates their recovery and reuse, improves stability and longevity, broadens applicability, and reduces overall processing and chemical conversion costs. Three-dimensional printing offers extraordinary flexibility for creating high-resolution complex structures that enable completely new reactor designs with versatile sub-micron functional features in macroscale objects. Immobilizing enzymes on or in 3D printed structures makes it possible to precisely control their spatial location for the optimal catalytic reaction. Combining the rapid advances in these two technologies is leading to completely new levels of control and precision in fabricating immobilized enzyme catalysts. The goal of this review is to promote further research by providing a critical discussion of 3D printed enzyme immobilization methods encompassing both post-printing immobilization and immobilization by physical entrapment during 3D printing. Especially, 3D printed gel matrix techniques offer mild single-step entrapment mechanisms that produce ideal environments for enzymes with high retention of catalytic function and unparalleled fabrication control. Examples from the literature, comparisons of the benefits and challenges of different combinations of the two technologies, novel approaches employed to enhance printed hydrogel physical properties, and an outlook on future directions are included to provide inspiration and insights for pursuing work in this promising field.

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An Immobilized Enzyme Reactor for Spatiotemporal Control over Reaction Products

Cited by 13 publications

References 57 publications

Using Microfluidics and Imaging SAMDI-MS To Characterize Reaction Kinetics

Using Microfluidics and Imaging SAMDI-MS To Characterize Reaction Kinetics

High-Throughput Enzyme Kinetics with 3D Microfluidics and Imaging SAMDI Mass Spectrometry

Advances in 3D Gel Printing for Enzyme Immobilization

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