Potentiometric aptasensing based on target-induced conformational switch of a DNA probe using a polymeric membrane...

In this article, we introduce a general, sensitive, facile, and label-free potentiometric assay based on metal-mediated DNA base pairs. A nucleic acid with one adenosine-5'-triphosphate (ATP) binding sequence (aptamer) in the middle and two cytosine(C)-rich sequences at the lateral portions was employed as a model. A rigid hairpin structure can be formed in the presence of Ag(+) ions, in which the C residues of the spatially separated nucleotides are linked by the ions. The strong interaction between Ag(+) ions and cytosines forms a stable C-Ag(+)-C structure, which could reduce the concentration of silver ions released from the polymeric membrane silver ion-selective electrode (ISE) at the sample-membrane interface and decrease the potential response. In the presence of its target, the aptamer (the loop sequence of the probe) binds specifically to the target via reaction incubation. Such target-binding induced aptamer conformational change prevents the formation of C-Ag(+)-C structure, leaving more silver ions at the sample-membrane interface, which can be detected by the silver ISE. ATP can be quantified in the range of 0.5-3.0 μM with a detection limit of 0.37 μM. The relative standard deviation for 5 μM ATP is 5.5%. For the proposed method, the combination of using ion fluxes of silver ions as modulating reagents and as signal reporters greatly simplifies the detection procedures. In addition, by changing the binding sequence in the middle of the probe, the present detection method will be able to explore new applications of ISE for the detection of a large variety of targets.