A versatile gold leaf immunosensor with a novel surface functionalization strategy based on protein L and trastuzumab for HER2 detection.

Although various sensors specifically developed for target analytes are available, affordable biosensing solutions with broad applicability are limited. In this study, a cost-effective biosensor for detecting human epidermal growth factor receptor 2 (HER2) was developed using custom-made gold leaf electrodes (GLEs). A novel strategy for antibody immobilization on a gold surface, for the first time mediated by protein L and HER2-specific antibody trastuzumab, was examined using commercial screen-printed gold electrodes and GLEs. A self-assembled monolayer of 11-mercaptoundecanoic acid (MUA) was formed on the gold surface, which was used to covalently immobilize protein L. Further binding of trastuzumab to the protein L was employed and HER2 detection was achieved through electrochemical impedance spectroscopy (EIS). The HER2 detection was examined in phosphate-buffered saline (PBS) and supplemented cell culture medium. The modified GLEs showed good specificity and high sensitivity of HER2 detection without any enrichment steps, achieving a limit of detection (LOD) of 1 ng mL in PBS and 2.7 ng mL in cell culture medium, making the proposed immunosensor a cost-effective and sensitive solution for detection in complex biological matrices.