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.
Original language | Undefined/Unknown |
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Pages (from-to) | 9781–6 |
Journal | Analytical Chemistry |
Volume | 80 |
Issue number | 24 |
DOIs | |
Publication status | Published - 2008 |
MoE publication type | A1 Journal article-refereed |
Keywords
- biosensor
- protein adsorption
- Fluorescence resonance energy transfer (FRET)
- Time-resolved fluorescence