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Issue 4, 2015
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Low-volume multiplexed proteolytic activity assay and inhibitor analysis through a pico-injector array

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Abstract

Secreted active proteases, from families of enzymes such as matrix metalloproteinases (MMPs) and ADAMs (a disintegrin and metalloproteinases), participate in diverse pathological processes. To simultaneously measure multiple specific protease activities, a series of parallel enzyme reactions combined with a series of inhibitor analyses for proteolytic activity matrix analysis (PrAMA) are essential but limited due to the sample quantity requirements and the complexity of performing multiple reactions. To address these issues, we developed a pico-injector array to generate 72 different reactions in picoliter-volume droplets by controlling the sequence of combinational injections, which allowed simultaneous recording of a wide range of multiple enzyme reactions and measurement of inhibitor effects using small sample volumes (~10 μL). Multiple MMP activities were simultaneously determined by 9 different substrates and 2 inhibitors using injections from a pico-injector array. Due to the advantages of inhibitor analysis, the MMP/ADAM activities of MDA-MB-231, a breast cancer cell line, were characterized with high MMP-2, MMP-3 and ADAM-10 activity. This platform could be customized for a wide range of applications that also require multiple reactions with inhibitor analysis to enhance the sensitivity by encapsulating different chemical sensors.

Graphical abstract: Low-volume multiplexed proteolytic activity assay and inhibitor analysis through a pico-injector array

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Publication details

The article was received on 02 Oct 2014, accepted on 23 Dec 2014 and first published on 23 Dec 2014


Article type: Paper
DOI: 10.1039/C4LC01162G
Citation: Lab Chip, 2015,15, 1153-1159
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    Low-volume multiplexed proteolytic activity assay and inhibitor analysis through a pico-injector array

    E. X. Ng, M. A. Miller, T. Jing, D. A. Lauffenburger and C. Chen, Lab Chip, 2015, 15, 1153
    DOI: 10.1039/C4LC01162G

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