A new electrochemical aptasensor based on a dual-signaling strategy and supersandwich assay

In this study, we develop a new electrochemical aptasensor by coupling two amplification strategies, including a dual signaling strategy and a supersandwich assay. In order to fabricate this aptasensor, a thiolated capture probe (CP) was first self-assembled on the gold electrode surface by Au-S bonds. After the addition of methylene blue (MB) modified signal probe 1 (SP1) and ferrocene (Fc) labeled signal probe 2 (SP2), supersandwich structure DNA, including multiple units of SP1 and SP2, was grown from the CP on the electrode surface. In the presence of ATP, the strong interaction between ATP and its aptamer (CP, SP1) leads to the disassembly of the supersandwich structure and thereby, the release of SP1 and SP2 from the gold electrode surface, resulting in a decrease of the MB and Fc signals. Taking "Signal gainMB + Signal gainFc" as the response signal, ATP can be detected sensitively; the detection limit is 2.1 nM, which is lower than that using either a single-signaling strategy or a traditional sandwich assay alone. Moreover, the new aptasensor also exhibits excellent specificity, selectivity, reliability and applicability. We believe that this new strategy will be helpful for fabricating sensitive and selective electrochemical aptasensors of other biomolecules and small molecules.