Highly Sensitive Reduced Graphene Oxide Impedance Sensor Harnessing π-Stacking Interaction Mediated Direct Deposition of Protein Probes

Graphene-based electrochemical impedance sensors have recently received much attention due to their outstanding sensing capability and economic viability. In this study, we present a novel means of constructing an impedance sensing platform via harnessing intrinsic pi-stacking interactions between probe protein molecules and reduced graphene oxide (RGO) substrate, obviating the need for introducing external chemical groups often required for covalent anchoring of the probes. To achieve this goal, protein molecules used as a probe were denatured to render their hydrophobic residues exposed in order to facilitate their direct pi-stacking interactions with the surface of RGO nanosheets. The protein molecules in denatured form, which would otherwise have difficulty in undergoing pi-stacking interactions with the RGO surface, were found to uniformly cover the RGO nanosheets at high density, conducive to providing a graphene-based impedance sensing platform capable of detecting a probe-specific analyte at high sensitivity. The proof-of-concept performance of thus-constructed RGO-based impedance sensors was demonstrated via selective detection of biological binding events of antigen antibody reaction at a femtomolar range. Notably, since the pi-stacking interaction can occur on the entire RGO surface, it can desirably exclude a backfill process indispensable for the conventional biosensors to suppress background noise signals. Since the procedure of pi-stacking mediated direct deposition of on-purpose denatured protein probes onto the RGO surface is facile and straightforward, the proposed strategy is anticipated to extend its applicability for fabrication of high performance graphene-based bio or chemical sensors.