Highly sensitive detection of thrombin using SERS-based magnetic aptasensors

Abstract

This paper reports a method of highly sensitive detection of thrombin using a surface-enhanced Raman scattering (SERS)-based magnetic aptasensor. Magnetic beads and gold nanoparticles (Au NPs) were used as supporting substrates and sensing probes, respectively. For this purpose, 15-mer thrombin-binding aptamers (TBA(15)) were immobilized onto the surface of magnetic beads, and then thrombin antigens and 29-mer thrombin-binding aptamer (TBA(29))-conjugated Au NPs were sequentially added for the formation of sandwich aptamer complexes. Quantitative analysis was performed by monitoring the intensity variation of a characteristic SERS signal of Raman reporter molecules. Because all of the reactions occur in solution, this SERS-based immunoassay technique can solve the diffusion-limited kinetic problems on a solid substrate. The limit of detection (LOD) of thrombin, determined by the SERS-based aptasensor, was estimated to be 0.27 pM. The proposed method is expected to be a good clinical tool for the diagnosis of a thrombotic disease. (C) 2013 Elsevier B.V. All rights reserved.