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    Quantum-Dot-Based Förster Resonance Energy Transfer Immunoassay for Sensitive Clinical Diagnostics of Low-Volume Serum Samples
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    † Institut d’Electronique Fondamentale, Université Paris-Sud, 91405 Orsay Cedex, France
    ‡ eBioscience, Inc., 10255 Science Center Drive, San Diego, California 92121, United States
    *Address correspondence to [email protected]
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    ACS Nano

    Cite this: ACS Nano 2013, 7, 8, 7411–7419
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    https://doi.org/10.1021/nn403253y
    Published August 2, 2013
    Copyright © 2013 American Chemical Society
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    Abstract

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    A myriad of quantum dot (QD) biosensor examples have emerged from the literature over the past decade, but despite their photophysical advantages, QDs have yet to find acceptance as standard fluorescent reagents in clinical diagnostics. Lack of reproducible, stable, and robust immunoassays using easily prepared QD-antibody conjugates has historically plagued this field, preventing researchers from advancing the deeper issues concerning assay sensitivity and clinically relevant detection limits on low-volume serum samples. Here we demonstrate a ratiometric multiplexable FRET immunoassay using Tb donors and QD acceptors, which overcomes all the aforementioned limitations toward application in clinical diagnostics. We demonstrate the determination of prostate specific antigen (PSA) in 50 μL serum samples with subnanomolar (1.6 ng/mL) detection limits using time-gated detection and two different QD colors. This concentration is well below the clinical cutoff value of PSA, which demonstrates the possibility of direct integration into real-life in vitro diagnostics. The application of IgG, F(ab′)2, and F(ab) antibodies makes our homogeneous immunoassay highly flexible and ready-to-use for the sensitive and specific homogeneous detection of many different biomarkers.

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    Supporting Information

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    PL decay curves of the following FRET immunoassays: (1) the “(Tb-IgG)-PSA-(QD650-IgG)”, (2) “(Tb-IgG)-PSA-(QD650-F(ab′)2)”, (3) “(Tb-IgG)-PSA-(QD650-F(ab))”, (4) “(Tb-F(ab))-PSA-(QD650-F(ab))”, and (5) “(Tb-IgG)-PSA-(QD605-IgG)”. This material is available free of charge via the Internet at http://pubs.acs.org.

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