Aptamer biosensor for label-free impedance spectroscopy detection of thrombin based on gold nanoparticles

This paper presents a simple electrochemical impedance spectroscopy (EIS) aptasensor based on an antithrombin-aptamer as a molecular recognition element. Improvement in sensitivity was achieved by utilizing gold nanoparticles (AuNPs), which were self-assembled on the surface of a bare electrode by using 1,6-Hexanedithiol as a medium. To quantify the amount of thrombin, changes in the interfacial electron transfer resistance (R-et) of the aptasensor were monitored using the redox couple of an [Fe(CN)(6)](3-/4-) probe. The plot of (R-eti - R-et0)/R-et0 against the logarithm of thrombin concentration is linear with over the range from 0.1 nM to 30 nM with a detection limit of 0.013 nM. Meanwhile, the packing density of aptamers was determined by cyclic voltammetric (CV) studies of redox cations (e.g., [Ru(NH3)(6)](3+)) which were electrostatically bound to the DNA phosphate backbones. The results indicate that the total amount of aptamer probes immobilized on the gold nanoparticle surface is sixfold higher than that on the bare electrode. The aptasensor also showed good selectivity for thrombin without being affected by the presence of other proteins. (C) 2011 Elsevier B.V. All rights reserved.