DNA sensor by using electrochemiluminescence of acridinium ester initiated by tripropylamine
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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.
KeywordsElectrochemiluminescence DNA sensor Acridinium ester Gold nanoparticles
This work was supported by the National Natural Science Foundation of P. R. China (grant nos. 20625517 and 20573101) and the Overseas Outstanding Young Scientist Program of the Chinese Academy of Sciences.
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