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An off–on–off electrochemiluminescence approach for ultrasensitive detection of thrombin

Deng, Li, Du, Ying, Xu, Jing-Juan, Chen, Hong-Yuan
Biosensors & bioelectronics 2014 v.59 pp. 58-63
biosensors, complementary DNA, detection limit, electrochemiluminescence, energy transfer, glassy carbon electrode, nanocrystals, nanogold, oligonucleotides, surface plasmon resonance, thrombin
This work demonstrates an aptasensor for ultrasensitive electrochemiluminescence (ECL) detection of thrombin based on an “off–on–off” approach. The system is composed of an Eu3+-doped CdS nanocrystals (CdS:Eu NCs) film on glassy carbon electrode (GCE) as ECL emitter. Then gold nanoparticles (AuNPs) labeled hairpin-DNA probe (ssDNA1) containing thrombin-binding aptamer (TBA) sequence was linked on the NCs film, which led to ECL quenching (off) as a result of Förster-resonance energy transfer (FRET) between the CdS:Eu NC film and the proximal AuNPs. Upon the occurrence of hybridization with its complementary DNA (ssDNA2), an ECL enhancement (on) occurred owing to the interactions of the excited CdS:Eu NCs with ECL-induced surface plasmon resonance (SPR) in AuNPs at large separation. Thrombin could induce ssDNA1 forming a G-quadruplex and cause the AuNPs to be close to CdS:Eu NCs film again, which resulted in an enhanced ECL quenching (off). This “off–on–off” system showed a maximum 7.4-fold change of ECL intensity due to the configuration transformation of ssDNA1 and provides great sensitivity for detection of thrombin in a wide detection range from 50aM to 1pM.