Aptamer biosensor based on fluorescence resonance energy transfer from upconverting phosphors to carbon nanoparticles for thrombin detection in human plasma

Wang, Y, Bao, L, Liu, Z and Pang, D 2011, 'Aptamer biosensor based on fluorescence resonance energy transfer from upconverting phosphors to carbon nanoparticles for thrombin detection in human plasma', Analytical Chemistry, vol. 83, no. 21, pp. 8130-8137.


Document type: Journal Article
Collection: Journal Articles

Title Aptamer biosensor based on fluorescence resonance energy transfer from upconverting phosphors to carbon nanoparticles for thrombin detection in human plasma
Author(s) Wang, Y
Bao, L
Liu, Z
Pang, D
Year 2011
Journal name Analytical Chemistry
Volume number 83
Issue number 21
Start page 8130
End page 8137
Total pages 8
Publisher American Chemical Society
Abstract We presented a new aptamer biosensor for thrombin in this work, which was based on fluorescence resonance energy transfer (FRET) from upconverting phosphors (UCPs) to carbon nanoparticles (CNPs). The poly(acrylic acid) (PAA) functionalized UCPs were covalently tagged with a thrombin aptamer (5′-NH 2 -GGTTGGTGTGGTTGG-3′), which bound to the surface of CNPs through π-π stacking interaction. As a result, the energy donor and acceptor were taken into close proximity, leading to the quenching of fluorescence of UCPs. A maximum fluorescence quenching rate of 89% was acquired under optimized conditions. In the presence of thrombin, which induced the aptamer to form quadruplex structure, the π-π interaction was weakened, and thus, the acceptor was separated from the donor blocking the FRET process. The fluorescence of UCPs was therefore restored in a thrombin concentration-dependent manner, which built the foundation of thrombin quantification. The sensor provided a linear range from 0.5 to 20 nM for thrombin with a detection limit of 0.18 nM in an aqueous buffer. The same linear range was obtained in spiked human serum samples with a slightly higher detection limit (0.25 nM), demonstrating high robustness of the sensor in a complex biological sample matrix. As a practical application, the sensor was used to monitor thrombin level in human plasma with satisfactory results obtained. This is the first time that UCPs and CNPs were employed as a donor-acceptor pair to construct FRET-based biosensors, which utilized both the photophysical merits of UCPs and the superquenching ability of CNPs and thus afforded favorable analytical performances. This work also opened the opportunity to develop biosensors for other targets using this UCPs-CNPs system.
Subject Sensor Technology (Chemical aspects)
Nanomaterials
Keyword(s) Up-Conversion
2-Photon Excitation
Homogeneous Immunoassay
Quantum Dots
Sensing Biomolecules
Hybridization Assay
Magnetic Beads
Graphene Oxide
Sandwich Assay
Platform
DOI - identifier 10.1021/ac201631b
Copyright notice © 2011 American Chemical Society
ISSN 0003-2700
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