Aptamer-based electrochemical detection of protein using enzymatic silver deposition

In this work an electrochemical protein detection based on enzymatic silver deposition has been proposed and applied to the detection of thrombin. The target protein, thrombin, was first captured by thrombin binding thiolated aptamer self-assembled monolayers (SAMs) on the gold electrode surface, and then sandwiched with another biotinylated thrombin binding aptamer for the association of alkaline phosphatase (Av-ALP). The attached Av-ALP enzymatically converts the nonelectroactive substrate p-aminophenyl phosphate (p-APP) to p-aminophenol (p-AP) which can reduce silver ions in solution leading to deposition of the metal onto the electrode surface. Finally, linear sweep voltammetry (LSV) is used to detect the amount of deposited silver. The peak current during the anodic scan was found to reflect the amount of the target protein captured into the sandwich configuration. The proposed approach has been successfully implemented for the detection of thrombin in the range of 0.1 nM to 1 mu M. Therefore, the current work has demonstrated enzymatic silver deposition for detection at aptamer-modified electrode. (C) 2009 Elsevier Ltd. All rights reserved.