Energy transfer-based biosensing of protease activity measured using an electroluminescent platform

Kim E. Sapsford; Steven Sun; Jesse Francis; Yordan Kostov; Avraham Rasooly; Dorothy Farrell; Hedi Mattoussi; Igor L. Medintz

doi:10.1117/12.809301

19 February 2009 Energy transfer-based biosensing of protease activity measured using an electroluminescent platform

Kim E. Sapsford, Steven Sun, Jesse Francis, Yordan Kostov, Avraham Rasooly, Dorothy Farrell, Hedi Mattoussi, Igor L. Medintz

Author Affiliations +

Proceedings Volume 7167, Frontiers in Pathogen Detection: From Nanosensors to Systems; 71670X (2009) https://doi.org/10.1117/12.809301
Event: SPIE BiOS, 2009, San Jose, California, United States

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.

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Kim E. Sapsford, Steven Sun, Jesse Francis, Yordan Kostov, Avraham Rasooly, Dorothy Farrell, Hedi Mattoussi, and 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); https://doi.org/10.1117/12.809301

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KEYWORDS

Fluorescence resonance energy transfer

Sensors

Luminescence

Biosensing

Charge-coupled devices

Electroluminescence

Quantum dots

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