Journal
BIOSENSORS & BIOELECTRONICS
Volume 43, Issue -, Pages 200-204Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2012.11.027
Keywords
Adenosine triphosphate; Aptasensor; Electrochemiluminescence; Probe intercalation; Tetrahedron-structured DNA
Categories
Funding
- NSF of China [21075068]
- National Basic Research Program of China (973 Program) [2011CB707703]
- Tianjin Natural Science Foundation [11JCZDJC22200]
- Fundamental Research Funds for the Central Universities
- NSF of China [21075068]
- National Basic Research Program of China (973 Program) [2011CB707703]
- Tianjin Natural Science Foundation [11JCZDJC22200]
- Fundamental Research Funds for the Central Universities
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Restricted target accessibility and surface-induced perturbation of the aptamer structure are the main limitations in single-stranded DNA aptamer-based electrochemical sensors. Chemical labeling of the aptamer with a probe at the end of aptamer is inefficient and time-consuming. In this work, tetrahedron-structured DNA (ts-DNA) and a functionalized oligonucleotide (FO) were used to develop an electrochemiluminescence (ECL) aptasensor with adenosine triphosphate (ATP) as a model target. The ts-DNA was formed with three thiolated oligonucleotides and one oligonucleotide containing anti-ATP aptamer. The FO contained a complementary strand to the anti-ATP aptamer and an intermolecular duplex for Ru(phen)(3)(2+) intercalation. After the ts-DNA was immobilized on the electrode surface through gold thiol interactions, hybridization between the anti-ATP aptamer and its complementary strand introduced the intercalated Ru(phen)(3)(2+) to the electrode. ECL emission from Ru(phen)(3)(2+) was observed with tripropylamine as a co-reactant. Once ATP reacted with its aptamer, the aptamer-complimentary strand duplex dissociated and the intermolecular duplex containing Ru(phen)(3)(2+) was released. The difference in emission before and after reaction with ATP was used to quantify ATP with a detection limit of 0.2 nM. The ts-DNA increased the sensitivity compared to conventional methods, and the intercalation strategy avoided a complex chemical labeling procedure. (c) 2012 Elsevier B.V. All rights reserved.
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