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DNA sensor by using electrochemiluminescence of acridinium ester initiated by tripropylamine

He, Yi, Zhang, Hao, Chai, Ying, Cui, Hua
Analytical and bioanalytical chemistry 2011 v.399 no.10 pp. 3451-3458
DNA, detection limit, electrochemiluminescence, electrodes, free radicals, gold, nanogold, nucleic acid hybridization, sensors (equipment), streptavidin
It was found that tripropylamine (TPA) could be used as a coreactant to initiate the electrochemiluminescence (ECL) of acridinium NHS ester (AE NHS) labels attached to DNA. The radicals generated in the electro-oxidation process of TPA reacted with AE NHS to form the excited N-methylacridone, giving rise to light emission. The AE/TPA ECL system was for the first time used as the detection system for developing an ECL-based DNA sensor. In the protocol, streptavidin-modified gold nanoparticles were firstly immobilized onto a thiol-treated gold electrode. The streptavidin could specifically interact with the biontinylated capture DNA. Afterwards, the target DNA and the AE-labeled report DNA were conjugated onto the electrode step by step due to the hybridization reactions, and a sandwich-type sensor was fabricated. The ECL signals of the sensor were obtained under pulse potential condition in alkaline solution containing 50.0 mmol L⁻¹ TPA. Under optimized experimental conditions, the linear range of the DNA sensor for the determination of the target DNA was from 5.0 × 10⁻¹⁵ to 5.0 × 10⁻¹² mol L⁻¹. The detection limit (S/N = 3) was 3.0 × 10⁻¹⁵ mol L⁻¹. Moreover, the sensor could specifically recognize the target DNA against one base-pair mismatched sequences, two base-pair mismatched sequences, and the noncomplementary sequences. It is of great application potential in clinic analysis.