High-Throughput Enzyme Kinetics with 3D Microfluidics and Imaging SAMDI Mass Spectrometry
- 26 September 2018
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 90 (21), 13096-13103
- https://doi.org/10.1021/acs.analchem.8b04391
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 paper describes a microfluidic device that has three inputs and performs 2,592 distinct enzyme reactions using only 150 µL of reagent with quantitative characterization. This paper 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 (Km). This approach expands the scope of imaging mass spectrometry (IMS), microfluidic detection strategies, and the design of high-throughput reaction systems.Keywords
Funding Information
- Defense Threat Reduction Agency (HDTRA1-15-1- 0052)
- National Science Foundation (DGE-1324585)
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