High-Throughput Mutiplexed Protease Activity Measurement Using a Droplet Based Microfluidic Platform with Picoinjector
Name
Griffith_High-throughput.pdf
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1.78 MB
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Adobe PDF
Checksum (MD5)
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Author(s)
Chen, Chia-Hung
Miller, Miles Aaron
Sarkar, Aniruddh
Beste, Michael T.
Lauffenburger, Douglas A.
Griffith, Linda G.
Han, Jongyoon
Date Issued
October 2012
Journal
Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences
Citation
Chen, Chia-Hung, et al. "High-Throughput Mutiplexed Protease Activity Measurement Using a Droplet Based Microfluidic Platform with Picoinjector." The 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, October 28-November 1, 2012, Okinawa, Japan.
Version
Author's final manuscript
Abstract
In this study, we integrated several components, including a droplet generator, a pico-injector, and an analytical inference technique, Proteolytic Activity Matrix Analysis (PrAMA), to create a platform for assessing multiple specific protease activity assays with minimal liquid handling and sample-requirement for personal medicine analysis. The microfluidic platform enables the direct measurements of protease enzyme activity, which is more physiologically informative than the standard measurements to determine the enzyme concentration alone. By tracking hundreds of picoliter droplets containing biological samples mixed with unique FRET-based protease substrates and inhibitors, the assay simultaneously infers multiple specific protease activities with minimal (<20µl) physiological sample requirement. We applied this method to in vitro study of an immortalized cell line established from a peritoneal endometriotic biopsy to ascertain the proteolytic activity response of TNF-α treatments.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Research Laboratory of Electronics
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Creative Commons Attribution-Noncommercial-Share Alike
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DOI of Published Version
http://www.rsc.org/images/loc/2012/pdf/W.1.22.pdf
