Sensitive Quantitative Protein Concentration Method Using Luminescent Resonance Energy Transfer on a Layer-by-Layer Europium(III) Chelate Particle Sensor



: Harma H, Dahne L, Pihlasalo S, Suojanen J, Peltonen J, Hanninen P

Publisher: American Chemical Society ACS

: 2008

Analytical Chemistry

ANALYTICAL CHEMISTRY

: ANAL CHEM

: 80

: 24

: 9781

: 9786

: 6

: 0003-2700

: 1520-6882

DOI: https://doi.org/10.1021/ac801960c


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.



Last updated on 2025-14-10 at 10:10