Multilevel electrochemical signal detections of metalloprotein heterolayers for bioelectronic device

In the present study, we investigated the simultaneous detection of multilevel electrochemical signals from various metalloprotein heterolayers for the bioelectronic devices. A layer-by-layer assembly method based on simple electrostatic interaction was introduced to form protein bilayers. The gold substrate was modified with poly (ethylene glycol) thiol acid as the precursor, which introduced negative charges to the surface. Based on the isoelectric point, net-charge controlled metalloproteins by pH adjustment were sequentially immobilized on this negatively charged substrate. The degree of protein immobilization on the gold substrate was confirmed by surface plasmon resonance spectroscopy, and the surface topology changes due to the protein immobilization were confirmed by atomic force microscopy. Redox signals in the protein layers were measured by cyclic voltammetry. As a result, various redox signals generated from different metalloproteins on a single electrode were monitored. This proposed method for the detection of multi-level electrochemical signals can be directly applied to bioelectronic devices that store multi-information in a single electrode. - Highlights: • We fabricated heterolayers composed of various metalloproteins. • Metalloproteins were immobilized by layer-by-layer assembly. • The degree of immobilization was controlled by the net charge of metalloproteins. • Various redox signals generated from heterolayers were well monitored