Plasmon field-enhanced fluorescence energy transfer for loop aptamer assay readout

Khulan Sergelen, Stefan Fossati, Aysegül Turupcu, Chris Oostenbrink, Bo Gunnar Liedberg, Wolfgang Knoll, Jakub Dostalek

    Publikation: Beitrag in FachzeitschriftArtikelBegutachtung

    Abstract

    Surface plasmon field-enhanced fluorescence energy transfer is employed for sensitive optical readout of a reversible hairpin aptamer assay that is suitable for continuous monitoring of low-molecular-weight chemical analytes. A hairpin aptamer specific to adenosine and adenosine triphosphate with Alexa Fluor 647 fluorophore attached to its 5′ end was anchored via 3′ end thiol to a gold thin film. Molecular spacers were used to control the distance of the fluorophore from the surface in the aptamer "off" and "on" states. The specific binding of the target analyte changes the aptamer conformation, which alters the distance of the fluorophore from the gold surface and translates to variations in the detected fluorescence intensity. The plasmonically mediated fluorescence signal increases the measured signal-to-noise ratio and allows for real-time observation of the analyte binding. Theoretical as well as experimental study of the optical signal dependence on fluorophore orientation, design of spacers, and angular distribution of collected light is presented for rational design of the assay. The detected sensor signal increased by a factor as large as 23 upon switching the aptamer from the "off" to "on" state due to the hairpin opening associated with the specific capture of target analyte.
    OriginalspracheEnglisch
    Seiten (von - bis)916-923
    Seitenumfang8
    FachzeitschriftACS Sensors
    Volume2
    Issue7
    DOIs
    PublikationsstatusVeröffentlicht - 2017

    Research Field

    • Biosensor Technologies

    Schlagwörter

    • adenosine triphosphate; aptamer; fluorescence energy transfer; plasmonic amplification; quenching; surface plasmon field-enhanced fluorescence

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