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dc.contributor.authorChopko Ahrens, Caroline
dc.contributor.authorChiswick, Evan
dc.contributor.authorRavindra, Kodihalli C.
dc.contributor.authorMiller, Miles Aaron
dc.contributor.authorRamseier, Julie Y.
dc.contributor.authorIsaacson, Keith B.
dc.contributor.authorLauffenburger, Douglas A
dc.contributor.authorGriffith, Linda G
dc.date.accessioned2020-06-22T15:40:33Z
dc.date.available2020-06-22T15:40:33Z
dc.date.issued2019-09
dc.date.submitted2019-08
dc.identifier.issn0006-2960
dc.identifier.issn1520-4995
dc.identifier.urihttps://hdl.handle.net/1721.1/125912
dc.description.abstractMetalloproteinases (MMPs) are zinc-dependent endopeptidases that cleave various proteins to regulate normal and diseased cellular functions, and as such, they play significant roles in human tissue development, homeostasis, and the pathogenesis of many diseases, including cancers, endometriosis, arthritis, etc. Most MMPs are produced as zymogenic latent enzymes that must be cleaved to activate their catalytic regions, and localized endogenous protein inhibitors further regulate activity. Accordingly, they operate within recursive networks to degrade extracellular matrix proteins and regulate cell signaling by cleaving growth factors and receptors at the cell surface and in the local pericellular environment. Thus, high-resolution information about the concentrations of specific active MMPs, revealing their intricate regulatory networks, may improve disease diagnosis and treatment. Here, we introduce a new and readily mastered method for measuring MMP activities in a multiplex fashion. We integrate aspects of activity-based enzyme labeling with commercial high-throughput, multiplexed protein quantification to yield the metalloproteinase activity multiplexed bead-based immunoassay (MAMBI). Assays of recombinant active MMP-1, -2, -3, -7, -8, -9, -12, and -13 establish the sensitivity and selectivity of MAMBI detection. Levels of active native MMPs are similarly characterized in conditioned cell culture medium, menstrual effluent, and uterine tissue. In a single MAMBI (5 μL), we achieve sensitivities equal to those from leading single-plex MMP activity detection strategies (e.g., 10-15 M for MMP-1). We also demonstrate high-throughput inhibitor screening via the MAMBI approach in complex, patient-derived samples.en_US
dc.description.sponsorshipNational Institutes of Health (Grant R01 EB010246)en_US
dc.language.isoen
dc.publisherAmerican Chemical Society (ACS)en_US
dc.relation.isversionofhttp://dx.doi.org/10.1021/acs.biochem.9b00584en_US
dc.rightsCreative Commons Attribution-Noncommercial-Share Alikeen_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/en_US
dc.sourceProf. Lauffenberger via Howard Silveren_US
dc.titleDevelopment and Application of the Metalloprotease Activity Multiplexed Bead-Based Immunoassay (MAMBI)en_US
dc.typeArticleen_US
dc.identifier.citationAhrens, Caroline C. et al. "Development and Application of the Metalloprotease Activity Multiplexed Bead-Based Immunoassay (MAMBI)." Biochemistry 58, 38 (September 2019): 3938–3942 © 2019 American Chemical Societyen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.relation.journalBiochemistryen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2020-06-19T13:31:58Z
dspace.date.submission2020-06-19T13:32:01Z
mit.journal.volume58en_US
mit.journal.issue38en_US
mit.licensePUBLISHER_CC


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