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

Harri Härmä, Lars Dähne, Sari Pihlasalo, Janne Suojanen, Jouko Peltonen, Pekka Hänninen

    Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

    25 Citeringar (Scopus)

    Sammanfattning

    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.
    OriginalspråkOdefinierat/okänt
    Sidor (från-till)9781–6
    TidskriftAnalytical Chemistry
    Volym80
    Utgåva24
    DOI
    StatusPublicerad - 2008
    MoE-publikationstypA1 Tidskriftsartikel-refererad

    Nyckelord

    • biosensor
    • protein adsorption
    • Fluorescence resonance energy transfer (FRET)
    • Time-resolved fluorescence

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