Direct Detection of DNA and DNA-Ligand Interaction by Impedance Spectroscopy

Abstract

In this work we present an impedimetric detection system for DNA-ligand interactions. The sensor system consists of thiol-modified single-stranded DNA chemisorbed to gold. Impedance measurements in the presence of the redox system ferri-/ferrocyanide show an increase in charge transfer resistance (Rct) after hybridisation of a complementary target. Different amounts of capture strands, used for gold electrode modification, result in surface coverages between 3 and 15 pmol/cm2 ssDNA. The relative change in Rct upon hybridisation increases with increasing amount of capture probe on the electrode from 1.5- to 4.5-fold. Impedimetric detection of binding events of a metal-intercalator ([Ru(phen)3]2+) and a groove binder (spermine) to double-stranded DNA is demonstrated. Binding of [Ru(phen)3]2+ and spermine exhibits a decrease in charge transfer resistance. Here, the ligand’s interaction leads to electrostatic shielding of the negatively charged DNA backbone. The impedance changes have been evaluated in dependence on the concentration of both DNA binders. Furthermore, the association of a single-stranded binding protein (SSBP) is found to cause an increase in charge transfer resistance only when incubated with single-stranded DNA. The specific binding of an anti-dsDNA antibody to the dsDNA-modified electrode surface decreases in contrast the interfacial impedance.