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An insertion approach electrochemical aptasensor for mucin 1 detection based on exonuclease-assisted target recycling.

Biosensors & bioelectronics (2015-04-17)
Wei Wen, Rong Hu, Ting Bao, Xiuhua Zhang, Shengfu Wang
RESUMEN

In this work, a sensitive exonuclease-assisted amplification electrochemical aptasensor through insertion approach was developed for the detection of mucin 1 (MUC 1). In order to construct the aptasensor, 6-Mercapto-1-hexanol (MCH) was used to block partial sites of gold electrode (GE), followed by thiolated capture probe self-assembled on GE. Methylene blue (MB) labeled aptamer hybridized with capture probe at both ends to form double-strand DNA. For the MB labeled termini was close to GE, the electrochemical response was remarkable. The presence of MUC 1 caused the dissociation of the double-strand DNA owing to the specific recognition of aptamer to MUC 1. Then exonuclease I (Exo I) selectively digested the aptamer which bound with MUC 1, the released MUC 1 participated new binding with the rest aptamer. Insertion approach improved the reproducibility and Exo I-catalyzed target recycling improved the sensitivity of the aptasensor significantly. Under optimal experimental conditions, the proposed aptasensor had a good linear correlation ranged from 10 pM to 1 μM with a detection limit of 4 pM (Signal to Noise ratio, S/N=3). The strategy had great potential for the simple and sensitive detection of other cancer markers.

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Sigma-Aldrich
6-Mercapto-1-hexanol, 97%
Sigma-Aldrich
1-Hexanethiol, 95%
Sigma-Aldrich
6-Mercapto-1-hexanol, 99%
Sigma-Aldrich
1-Hexanethiol, 99% (GC)