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Proceedings Paper

Energy transfer-based biosensing of protease activity measured using an electroluminescent platform
Author(s): Kim E. Sapsford; Steven Sun; Jesse Francis; Yordan Kostov; Avraham Rasooly; Dorothy Farrell; Hedi Mattoussi; Igor L. Medintz
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Paper Abstract

We present a biosensing platform that uses spatial electroluminescent (EL) illumination combined with charge-coupled device (CCD)-based detection for fluorescence measurements. The resulting EL-CCD detector platform was used to monitor different protease activities with substrates labeled for fluorescence resonance energy transfer (FRET)-based assays. The first uses a commercial FITC/DABCYL-SNAP-25 peptide substrate to monitor the activity of the light chain derivative (LcA) of botulinum neurotoxin A, achieving a limit of detection (LOD) of 1.25 nM (62 ng/ml). The second protease activity assay measured trypsin proteolysis using peptide substrates immobilized onto semiconductor quantum dot (QD) nanoparticles with a LOD of 6.2 nM trypsin (140 ng/ml). The specific ovomucoid inhibition of trypsin activity was also monitored. The highlighted studies clearly demonstrate the utility of the EL-CCD detector platform for monitoring fluorescent-based protease activity assays with potential healthcare applications, including point-of-care diagnostics.

Paper Details

Date Published: 19 February 2009
PDF: 10 pages
Proc. SPIE 7167, Frontiers in Pathogen Detection: From Nanosensors to Systems, 71670X (19 February 2009); doi: 10.1117/12.809301
Show Author Affiliations
Kim E. Sapsford, U.S. Food and Drug Administration (United States)
Steven Sun, U.S. Food and Drug Administration (United States)
Univ. of Maryland, Baltimore County (United States)
Jesse Francis, U.S. Food and Drug Administration (United States)
Univ. of Maryland, Baltimore County (United States)
Yordan Kostov, Univ. of Maryland, Baltimore County (United States)
Avraham Rasooly, U.S. Food and Drug Administration (United States)
National Cancer Institute (United States)
Dorothy Farrell, Naval Research Lab. (United States)
Hedi Mattoussi, Naval Research Lab. (United States)
Igor L. Medintz, Naval Research Lab. (United States)


Published in SPIE Proceedings Vol. 7167:
Frontiers in Pathogen Detection: From Nanosensors to Systems
Philippe M. Fauchet, Editor(s)

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