A1 Refereed original research article in a scientific journal
Sensitive Quantitative Protein Concentration Method Using Luminescent Resonance Energy Transfer on a Layer-by-Layer Europium(III) Chelate Particle Sensor
Authors: Harma H, Dahne L, Pihlasalo S, Suojanen J, Peltonen J, Hanninen P
Publisher: American Chemical Society ACS
Publication year: 2008
Journal:: Analytical Chemistry
Journal name in source: ANALYTICAL CHEMISTRY
Journal acronym: ANAL CHEM
Volume: 80
Issue: 24
First page : 9781
Last page: 9786
Number of pages: 6
ISSN: 0003-2700
eISSN: 1520-6882
DOI: https://doi.org/10.1021/ac801960c
Abstract
A particle-based protein quantification method was developed. The method relies on adsorption of proteins on particles and time-resolved fluorescence resonance energy transfer (TR-FRET). Layer-by-layer (LbL) particles containing europium(III) chelate donor were prepared. A protein labeled with an acceptor was adsorbed onto the particles and near-infrared energy transfer signal was detected in time-gated detection mode. Sample proteins efficiently occupied the particle surface preventing binding of the acceptor-labeled protein leading to a particle sensor with a significant signal change. We detected subnanomolar protein concentration using the rapid and simple mix-and-measure method with a coefficient of variation below 10%. Compared to known protein concentration methods, the developed method required no hazardous substances or elevated temperature to reach the high-sensitivity level.
A particle-based protein quantification method was developed. The method relies on adsorption of proteins on particles and time-resolved fluorescence resonance energy transfer (TR-FRET). Layer-by-layer (LbL) particles containing europium(III) chelate donor were prepared. A protein labeled with an acceptor was adsorbed onto the particles and near-infrared energy transfer signal was detected in time-gated detection mode. Sample proteins efficiently occupied the particle surface preventing binding of the acceptor-labeled protein leading to a particle sensor with a significant signal change. We detected subnanomolar protein concentration using the rapid and simple mix-and-measure method with a coefficient of variation below 10%. Compared to known protein concentration methods, the developed method required no hazardous substances or elevated temperature to reach the high-sensitivity level.
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