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Modulation of Intracellular Quantum Dot to Fluorescent Protein Förster Resonance Energy Transfer via Customized Ligands and Spatial Control of Donor–Acceptor Assembly
Open AccessArticle

Evaluating Quantum Dot Performance in Homogeneous FRET Immunoassays for Prostate Specific Antigen

1
NanoBioPhotonics, Institut d’Electronique Fondamentale, Université Paris-Saclay, Université Paris-Sud, CNRS, 91405 Orsay, France
2
Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, BC V6T 1Z1, Canada
*
Author to whom correspondence should be addressed.
Academic Editor: Alexander Star
Sensors 2016, 16(2), 197; https://doi.org/10.3390/s16020197
Received: 30 November 2015 / Revised: 11 January 2016 / Accepted: 2 February 2016 / Published: 4 February 2016
(This article belongs to the Special Issue FRET Biosensors)
The integration of semiconductor quantum dots (QDs) into homogeneous Förster resonance energy transfer (FRET) immunoassay kits for clinical diagnostics can provide significant advantages concerning multiplexing and sensitivity. Here we present a facile and functional QD-antibody conjugation method using three commercially available QDs with different photoluminescence (PL) maxima (605 nm, 655 nm, and 705 nm). The QD-antibody conjugates were successfully applied for FRET immunoassays against prostate specific antigen (PSA) in 50 µL serum samples using Lumi4-Tb (Tb) antibody conjugates as FRET donors and time-gated PL detection on a KRYPTOR clinical plate reader. Förster distance and Tb donor background PL were directly related to the analytical sensitivity for PSA, ...which resulted in the lowest limits of detection for Tb-QD705 (2 ng/mL), followed by Tb-QD655 (4 ng/mL), and Tb-QD605 (23 ng/mL). Duplexed PSA detection using the Tb-QD655 and Tb-QD705 FRET-pairs demonstrated the multiplexing ability of our immunoassays. Our results show that FRET based on QD acceptors is suitable for multiplexed and sensitive biomarker detection in clinical diagnostics. View Full-Text
Keywords: Qdots; antibodies; bioconjugation; terbium; FRET; time-gating; homogeneous; immunoassay; serum; TPSA; clinical diagnostics Qdots; antibodies; bioconjugation; terbium; FRET; time-gating; homogeneous; immunoassay; serum; TPSA; clinical diagnostics
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MDPI and ACS Style

Bhuckory, S.; Lefebvre, O.; Qiu, X.; Wegner, K.D.; Hildebrandt, N. Evaluating Quantum Dot Performance in Homogeneous FRET Immunoassays for Prostate Specific Antigen. Sensors 2016, 16, 197. https://doi.org/10.3390/s16020197

AMA Style

Bhuckory S, Lefebvre O, Qiu X, Wegner KD, Hildebrandt N. Evaluating Quantum Dot Performance in Homogeneous FRET Immunoassays for Prostate Specific Antigen. Sensors. 2016; 16(2):197. https://doi.org/10.3390/s16020197

Chicago/Turabian Style

Bhuckory, Shashi; Lefebvre, Olivier; Qiu, Xue; Wegner, Karl D.; Hildebrandt, Niko. 2016. "Evaluating Quantum Dot Performance in Homogeneous FRET Immunoassays for Prostate Specific Antigen" Sensors 16, no. 2: 197. https://doi.org/10.3390/s16020197

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