Convenient Colorimetric Detection of Thrombin via Aptamer-Mediated Inhibition and Restoration of the Oxidase Activity of Nanoceria

Abstract

Cerium oxide nanoparticles, also called nanoceria, have recently gained much attention as oxidase-mimicking nanozymes that catalyze the oxidation of chromogenic substrates for color generation without the addition of H2O2. Herein, we have developed a unique colorimetric biosensor for thrombin in human blood plasma, which relies on thrombin-binding aptamer (TBA)-mediated inhibition of the oxidase activity of nanoceria and its restoration by very selective interactions of TBA with target thrombin. In this system, nanoceria were first incubated with TBA, resulting in quick reduction of the oxidase activity of nanoceria via the adsorption of single-stranded (ss)DNA-type TBA on nanoceria. By the addition of sample solutions containing target thrombin, TBA bound on the nanoceria would strongly interact with free thrombin and be detached from the nanoceria, thereby increasing the available surface area of the nanoceria and consequently enhancing the oxidase activity. Using this strategy, target thrombin was successfully detected at concentrations as low as 100 pM over a wide linear range from 0.1 to 10 nM. The diagnostic capability of this method has been demonstrated by detecting thrombin in human blood plasma, showing its great potential in the practical applications.