An "off-on" electrochemiluminescence biosensor coupled with strand displacement-powered 3D micromolecule walking nanomachine for ultrasensitive detection of adenosine triphosphate.

A three-dimensional (3D) micromolecule walking nanomachine propelled by strand displacement was developed to establish a novel switching electrochemiluminescence (ECL) biosensor for ultrasensitive detection of adenosine triphosphate (ATP). Generally, walking nanomachines reported previously were limited to DNA walkers, while the proposed 3D walking nanomachine focused on the micromolecule walker. Firstly, TiO2 and silver nanoparticles (Ag NPs) functionalized N-(4-aminobutyl)-N-(ethylisoluminol) (Ag-ABEI) were deposited onto the electrode surface to offer an enhanced ECL signal, resulting from the double catalytic effect of TiO2 and Ag NPs for H2O2. Following, dopamine (DA)-labeled DNA duplex probes (S1/S2-DA) immobilized onto the modified electrode cut down the original ECL signal due to the quenching of DA toward ABEI (signal-off). Target ATP walker moved along the 3D DNA track, simultaneously releasing numerous DNA3, which was applied to displace S2-DA, resulting in the quenched ECL intensity recovery (signal-on). As a result, the biosensor showed a low limit of detection down to 0.5 nM (S/N = 3) and was successfully employed to determine ATP in human serum samples. Thus, the established biosensing strategy holds great potential for biochemical studies and clinical diagnosis.