An amplified electrochemical aptasensor for thrombin detection based on pseudobienzymic Fe3O4-Au nanocomposites and electroactive hemin/G-quadruplex as signal enhancers.

A sensitive and selective electrochemical aptasensor for thrombin detection was constructed based on hemin/G-quadruplex as the signal label and Fe3O4-Au nanocomposites with glucose oxidase (GOx-) and peroxide-mimicking enzyme activity as the signal enhancers. Due to their large surface area and good biocompatibility, Fe3O4-Au nanocomposites were employed to immobilize electroactive hemin/G-quadruplex, which was formed by the conjugation between a single-stranded guanine-rich nucleic acid and hemin. Based on the GOx-mimicking enzyme activity, Au nanoparticles on the surface of the Fe3O4-Au nanocomposites effectively catalyzed the oxidization of glucose in the presence of dissolved O2, accompanied by the production of H2O2. Both the Fe3O4 cores of Fe3O4-Au nanocomposites and hemin/G-quadruplex with H2O2-mimicking enzyme activity could catalyze the reduction of the generated H2O2, which promoted the electron transfer of hemin and amplified the electrochemical signal. The proposed electrochemical aptasensor had a wide dynamic linear range of 0.1 pM to 20 nM with a lower detection limit of 0.013 pM, which provided a promising method for a sensitive assay for the detection of proteins in electrochemical aptasensors.