Next Article in Journal
A Walk through Recent Nitro Chemistry Advances
Next Article in Special Issue
Lanthanide Luminescence in Visible-Light-Promoted Photochemical Reactions
Previous Article in Journal
Glycation and Oxidative Stress Increase Autoantibodies in the Elderly
Previous Article in Special Issue
Two Beta-Phosphorylamide Compounds as Ligands for Sm3+, Eu3+, and Tb3+: X-ray Crystallography and Luminescence Properties
Article

Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET

1
CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, 91198 Gif-sur-Yvette, France
2
Federal Institute for Materials Research and Testing (BAM), Division Biophotonics, Richard-Willstaetter-Strasse 11, 12489 Berlin, Germany
3
School of Medicine and Pharmacy, Ocean University of China. 5, Yushan Road, Qingdao 266003, Shandong, China
4
Thermo Fisher Scientific, 5781 Van Allen Way, Carlsbad, CA 92008, USA
5
Thermo Fisher Scientific Cezanne SAS, Clinical Diagnostic Division, 30000 Nimes, France
6
Laboratoire COBRA (Chimie Organique, Bioorganique, Réactivité et Analyse), Université de Rouen Normandie, CNRS, INSA, 76821 Mont-Saint-Aignan, France
*
Author to whom correspondence should be addressed.
Academic Editor: Eszter Borbas
Molecules 2020, 25(16), 3679; https://doi.org/10.3390/molecules25163679
Received: 13 July 2020 / Revised: 6 August 2020 / Accepted: 9 August 2020 / Published: 12 August 2020
(This article belongs to the Special Issue Luminescent Lanthanide Complexes)
Time-gated Förster resonance energy transfer (TG-FRET) between Tb complexes and luminescent semiconductor quantum dots (QDs) provides highly advantageous photophysical properties for multiplexed biosensing. Multiplexed Tb-to-QD FRET immunoassays possess a large potential for in vitro diagnostics, but their performance is often insufficient for their application under clinical conditions. Here, we developed a homogeneous TG-FRET immunoassay for the quantification of carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), and prostate-specific antigen (PSA) from a single serum sample by multiplexed Tb-to-QD FRET. Tb–IgG antibody donor conjugates were combined with compact QD-F(ab’)2 antibody acceptor conjugates with three different QDs emitting at 605, 650, and 705 nm. Upon antibody–antigen–antibody sandwich complex formation, the QD acceptors were sensitized via FRET from Tb, and the FRET ratios of QD and Tb TG luminescence intensities increased specifically with increasing antigen concentrations. Although limits of detection (LoDs: 3.6 ng/mL CEA, 3.5 ng/mL NSE, and 0.3 ng/mL PSA) for the triplexed assay were slightly higher compared to the single-antigen assays, they were still in a clinically relevant concentration range and could be quantified in 50 µL serum samples on a B·R·A·H·M·S KRYPTOR Compact PLUS clinical immunoassay plate reader. The simultaneous quantification of CEA, NSE, and PSA at different concentrations from the same serum sample demonstrated actual multiplexing Tb-to-QD FRET immunoassays and the potential of this technology for translation into clinical diagnostics. View Full-Text
Keywords: lanthanides; nanoparticles; fluorescence; biosensing; multiplexing; PSA; NSE; CEA lanthanides; nanoparticles; fluorescence; biosensing; multiplexing; PSA; NSE; CEA
Show Figures

Figure 1

MDPI and ACS Style

Bhuckory, S.; Wegner, K.D.; Qiu, X.; Wu, Y.-T.; Jennings, T.L.; Incamps, A.; Hildebrandt, N. Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET. Molecules 2020, 25, 3679. https://doi.org/10.3390/molecules25163679

AMA Style

Bhuckory S, Wegner KD, Qiu X, Wu Y-T, Jennings TL, Incamps A, Hildebrandt N. Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET. Molecules. 2020; 25(16):3679. https://doi.org/10.3390/molecules25163679

Chicago/Turabian Style

Bhuckory, Shashi, K. D. Wegner, Xue Qiu, Yu-Tang Wu, Travis L. Jennings, Anne Incamps, and Niko Hildebrandt. 2020. "Triplexed CEA-NSE-PSA Immunoassay Using Time-Gated Terbium-to-Quantum Dot FRET" Molecules 25, no. 16: 3679. https://doi.org/10.3390/molecules25163679

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop