Enzyme-amplified electrochemical detection of DNA using electrocatalysis of ferrocenyl-tethered dendrimer

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We have developed a sandwich-type enzyme-linked DNA sensor as a new electrochemical method to detect DNA hybridization. A partially ferrocenyl-tethered poly(amidoamine) dendrimer (Fc-D) was used as an electrocatalyst to enhance the electronic signals of DNA detection as well as a building block to immobilize capture probes. Fc-D was immobilized on a carboxylic acid-terminated self-assembled monolayer (SAM) by covalent coupling of unreacted amine in Fc-D to the acid. Thiolated capture probe was attached to the remaining amine groups of Fc-D on the SAM via a bifunctional linker. The target DNA was hybridized with the capture probe, and an extension in the DNA of the target was then hybridized with a biotinylated detection probe. Avidin-conjugated alkaline phosphatase was bound to the detection probe and allowed to generate the electroactive label, p-aminophenol, from p-aminophenyl phosphate enzymatically. p-Aminophenol diffuses into the Fc-D layer and is then electrocatalytically oxidized by the electronic mediation of the immobilized Fc-D, which leads to a great enhancement in signal. Consequently, the amount of hybridized target can be estimated using the intensity of electrocatalytic current. This DNA sensor exhibits a detection limit of 20 fmol. Our method was also successfully applied to the sequence-selective discrimination between perfectly matched and single-base mismatched target oligonucleotides.
Publisher
AMER CHEMICAL SOC
Issue Date
2003-11
Language
English
Article Type
Article
Keywords

QUARTZ-CRYSTAL-MICROBALANCE; SELF-ASSEMBLED MONOLAYERS; SURFACE-PLASMON RESONANCE; NUCLEIC-ACID; AMPEROMETRIC DETECTION; ALKALINE-PHOSPHATASE; MODIFIED ELECTRODE; IMPROVED SUBSTRATE; CARBON ELECTRODES; MIXED MONOLAYERS

Citation

ANALYTICAL CHEMISTRY, v.75, no.21, pp.5665 - 5672

ISSN
0003-2700
URI
http://hdl.handle.net/10203/19336
Appears in Collection
CH-Journal Papers(저널논문)
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