Photodriven Regeneration of G-Quadruplex Aptasensor for Sensitively Detecting Thrombin

Aptamers have been widely used as recognition elements in electrochemical sensors. However, as the most expensive consumable, the aptasensors regeneration is still a critical challenge for sustainable feasibility and attracting great interest from researchers, due to the high affinity between the aptamers and their targets (the dissociation constant Kd is low to subnanomolar or nanomolar). In this work, we propose a photochromic five-azobenzene-inserted thrombin-aptamer based aptasensor to improve the regenerativity. With ultraviolet light exposure, the trans-structure of azobenzene changes to cis-structure, and open the folded aptamer to realize the aptasensor regeneration. The limit of detection can be sensitive to 3 pM (S/N = 3). The thrombin concentrations were detected to be 2.48 ± 0.02 and 20.26 ± 0.98 nM (n = 3) in duck whole blood and blood serum, respectively. Utilizing surface plasmon resonance, we demonstrated that the certain azobenzene moieties can exactly increase Kd of aptamer-thrombin bounding. The photodriven conversion of thrombin-aptamer from G-quadruplex to loosen structure approaches a convenient regeneration for aptasensor, which will promote its popularization and sustainable feasibility.