Deaeration of incubation buffer to minimize electrochemical impedance spectroscopy signal drift on screen-printed gold electrodes and its use for label-free detection of vancomycin

Disposable screen-printed gold electrodes (SPAuEs) are used for the development of numerous immunosensors owing to their ease of modification with biorecognition molecules. A major limitation in the development of SPAuEs based biosensors is the single-use of the electrodes leading to higher costs for sensor development, whereas multiple measurements can affect the sensor stability and reproducibility. Especially, label-free electrochemical impedance spectroscopy (EIS) based SPAuEs sensors are prone to signal drift resulting in false-positive or false-negative data questioning the reliability of the assay. We found that repeated EIS measurements on SPAuEs in ferri- and ferrocyanide solution yielded a significant increase in resistance to charge transfer (Rct) value (positive signal drift) even in a control solution (without the target analyte). The positive signal drift follows the typical pattern of a concentration-dependent calibration curve, rendering repeated measurement error-prone for sensing purposes. In this study, it is shown that the incubation of the SPAuEs in nitrogen purged deaerated phosphate buffered saline (N-PBS) minimized this signal drift in repeated/multiple EIS measurements. This method was then used to develop a peptide-based EIS biosensor for the detection of Vancomycin, a last-line antibiotic used for the treatment of severe multidrug-resistant bacterial infection. This protocol can be followed for the biosensing of other clinically important biomarkers.