NDLI: Ultrasensitive aptamer-based thrombin assay based on metal enhanced fluorescence resonance energy transfer

Content Provider	Springer Nature Link
Author	Sui, Ning Wang, Lina Xie, Fengxia Liu, Fengya Xiao, Hailian Liu, Manhong Yu, William W.
Copyright Year	2016
Abstract	A sensitive “on-off” fluorescent protocol for thrombin detection is demonstrated. Firstly, thrombin aptamers which hybridize with labeled help DNA were immobilized on the surface of Ag@SiO2 nanoparticles (NPs). The silver core causes the label Cy5 to display strong metal-enhanced fluorescence. On addition of thrombin and graphene oxide (GO), thrombin (with its high affinity for the aptamers) displaces the Cy5-labeled help DNA which then binds to the surface of GO via π-stacking, causing fluorescence quenching of Cy5. The findings were used to design a thrombin assay that has a 0.05 nM detection limit and excellent selectivity. It was applied to the quantification of thrombin in spiked serum samples where is showed recoveries ranging from 97 % to 107 %, with relative standard deviations between 2.2 and 4.5 %. Graphical abstract A sandwich architecture of type Ag@SiO2-DNA-Cy5 was fabricated. The fluorescence of Cy5 is enhanced by Ag@SiO2, and quenched by GO. Upon the introduction of thrombin and GO, (a) the quadruplex-thrombin complex is formed, (b) the DNA duplexes are dismissed, and (c) helper DNA is released, which then is absorbed by GO, this leading to quenching.
Starting Page	1563
Ending Page	1570
Page Count	8
File Format	PDF
ISSN	00263672
Journal	Microchimica Acta
Volume Number	183
Issue Number	5
e-ISSN	14365073
Language	English
Publisher	Springer Vienna
Publisher Date	2016-02-25
Publisher Place	Vienna
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	FRET Ag@SiO2 Transmission electron microscopy Graphene oxide Bioconjugation Help DNA Nanochemistry Nanotechnology Characterization and Evaluation of Materials Analytical Chemistry Microengineering
Content Type	Text
Resource Type	Article
Subject	Analytical Chemistry

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