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

Abstract

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.

Key concepts in muscle regeneration: muscle “cellular ecology” integrates a gestalt of cellular cross-talk, motility, and activity to remodel structure and restore function

“…Physical activity likely influences the resolution of inflammation, where the timing is critical; myotube alignment during regeneration; and the role of the ECM and ECM-bound ligands in guiding cells during movement. Myoblast migration (Roveimiab et al 2020 ) and the mechanisms of axonal targeting to neuromuscular junctions through synchronized myogenesis and nerve–muscle connectivity during regeneration (Daneshvar et al 2020 ) can be closely scrutinized by microfluidics applications, now used for high-throughput screening for mutations (Markin et al 2021 ) and biological assays (Grant et al 2018 ).…”

Using Microfluidics and Imaging SAMDI-MS To Characterize Reaction Kinetics

“…After flowing the reactants through the device, we removed the PDMS block from the chip and applied matrix to the chip as previously described. 14…”

The Present and Future Role of Microfluidics for Protein and Peptide-Based Therapeutics and Diagnostics

“…Measuring enzyme activity using an MF platform is not necessarily a contemporary idea, as many MF designs are centered around this [71,72]. However, due to recent ad-…”

“…Measuring enzyme activity using an MF platform is not necessarily a contemporary idea, as many MF designs are centered around this [71,72]. However, due to recent advances in neighboring fields, namely additive manufacturing (AM) and nano-detection, the capacity for MFs to act as an area-leading platform has been propelled.…”