19 February 2009 Energy transfer-based biosensing of protease activity measured using an electroluminescent platform
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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.
© (2009) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Kim E. Sapsford, Kim E. Sapsford, Steven Sun, Steven Sun, Jesse Francis, Jesse Francis, Yordan Kostov, Yordan Kostov, Avraham Rasooly, Avraham Rasooly, Dorothy Farrell, Dorothy Farrell, Hedi Mattoussi, Hedi Mattoussi, Igor L. Medintz, Igor L. Medintz, } "Energy transfer-based biosensing of protease activity measured using an electroluminescent platform", Proc. SPIE 7167, Frontiers in Pathogen Detection: From Nanosensors to Systems, 71670X (19 February 2009); doi: 10.1117/12.809301; https://doi.org/10.1117/12.809301
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