Greatly extended storage stability of electrochemical DNA biosensors using ternary thiolated self-assembled monolayers.

While high storage stability of sequence-selective DNA biosensors is crucial towards their routine applications, commonly used electrochemical hybridization biosensors are characterized with limited storage stability. In this article we demonstrate that recently developed ternary thiolated monolayers impart dramatic improvement in the storage stability of DNA electrochemical biosensors. In particular, highly stable multicomponent interfaces are prepared by co-immobilizing the thiolated capture probe (SHCP) with 1,6-hexanedithiol (HDT) on gold substrates, followed by the incorporation of 6-mercapto-1-hexanol (MCH) diluent. The resulting (SHCP/HDT+MCH) DNA hybridization recognition platform offers substantially higher storage stability compared to conventional binary (SHCP+MCH) monolayers. The (SHCP/HDT+MCH) ternary monolayers maintain their initial signal (S)-to-noise (N) ratio (S/N) over a prolonged 3 months period upon storage at 4 °C, compared to the rapid sensitivity loss observed using the common binary interfaces. This attractive stability performance promises the convenient usage of pre-prepared electrodes after prolonged time storage without any treatment. Such dramatic improvements in the storage stability have been achieved through a rational optimization of the concentration ratio of the SHCP and the other components of the ternary SAM. The improved storage stability of SHCP/HDT+MCH interfaces observed at higher concentrations of SHCP is attributed to a hindered displacement of SHCP by MCH in the resulting compact layers. The ability to design highly stable nucleic acid interfaces using common chemicals obviates the need of using specialized expensive reagents.