Fabrication of reusable electrochemical biosensor and its application for the assay of α-glucosidase activity

A reusable biosensor has been fabricated in this work for the assay of α-glucosidase activity and the inhibitor screening. In this design, the aptamer of ATP is split as split aptamer 1 (Apt 1) and split aptamer 2 (Apt 2), and Apt 2 can link gold nanoparticles (AuNPs) modified with Apt 1 and 4-aminophenyl-α-d-glucopyranoside (pAPG). Consequently, the functional AuNPs can be immobilized onto the surface of gold electrode, allowing for salt-induced regeneration. In the presence of α-glucosidase, the glycosyl of pAPG is cut off, and the electroactive phenolic hydroxyls appear to give a strong current signal. Furthermore, the biosensor can be recovered very easily by incubating it in water to dissociate the AuNPs modified with Apt 1 and pAPG. So, a new biosensor for α-glucosidase activity detection and inhibitor screening is developed based on enzyme-activated signal generation and recovery. The biosensor may also exhibit good sensitivity for α-glucosidase determination with the detection limit 0.005 U/mL and can be reused by water-washing regeneration with good repeatability. Meanwhile this biosensor can also be utilized for inhibitor screening, which may have potential for clinical applications.