Journal
BIOSENSORS & BIOELECTRONICS
Volume 26, Issue 5, Pages 2645-2650Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2010.11.028
Keywords
Aptamer biosensor; ECL aptamer biosensor; Junction-probe; Thrombin; Electrochemiluminescence
Categories
Funding
- NSFC [40875087]
- Ministry of Health P. R China [200902009]
- National Basic Research Program of China [2010CB732403]
- Fujian Provincial Department of Science and Technology [2009J01023, 2010H6010]
- Education Department of Fujian Province [JA10295]
- Fujian Health Department [2010126]
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A novel signal-on junction-probe electrogenerated chemiluminescence (ECL) aptamer biosensor has been developed for the detection of ultratrace thrombin based on a structure-switching ECL-quenching mechanism. The ECL aptamer biosensor comprises two main parts: an ECL substrate and an ECL intensity switch. The ECL substrate was made by modifying the complex of Au nanoparticle and ruthenium (II) tris-bipyridine (Ru(bpy)(3)(2+)-AuNPs) on the surface of gold electrode (GE), and the ECL intensity switch contains three probes designed according to the junction-probe strategy. The first probe is capture probe (Cp) which was functionalized with a thiol group at one end and covalently attached to Ru(bpy)(3)(2+)-AuNPs modified GE through S-Au bonding. The second probe is aptamer probe (Ap), which containing 15-base anti-thrombin DNA aptamer. The third one is ferrocene-labeled probe (Fp), which was functionalized with ferrocene tag at one end. We demonstrated that, in the absence of thrombin, Cp, Ap and Fp will hybridize to form a ternary Y junction structure and resulted in a quenching of ECL of Ru(bpy)(3)(2+). Whereas, in the presence of thrombin, the Ap prefers to form the G-quadruplex aptamer-thrombin complex and lead to an obvious recovery of ECL of Ru(bpy)(3)(2+), which provided a sensing platform for the detection of thrombin. Using this reusable sensing platform, a simple, rapid and selective signal-on ECL aptamer biosensor for the detection of thrombin with a detection limit of 8.0 x 10(-15) M has been developed. The success in the present biosensor served as a significant step towards the development of monitoring ultratrace thrombin in clinical detection. (C) 2010 Elsevier B.V. All rights reserved.
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