Biofunctionalization of Graphene‐Based FET Sensors through Heterobifunctional Nanoscaffolds: Technology Validation toward Rapid COVID‐19 Diagnostics and Monitoring

The biofunctionalization of graphene field‐effect transistors (GFETs) through vinylsulfonated‐polyethyleneimine nanoscaffold is presented for enhanced biosensing of severe acute respiratory‐related coronavirus 2 (SARS‐CoV‐2) spike protein and human ferritin, two targets of great importance for the rapid diagnostic and monitoring of individuals with COVID‐19. The heterobifunctional nanoscaffold enables covalent immobilization of binding proteins and antifouling polymers while the whole architecture is attached to graphene by multivalent π–π interactions. First, to optimize the sensing platform, concanavalin A is employed for glycoprotein detection. Then, monoclonal antibodies specific against SARS‐CoV‐2 spike protein and human ferritin are anchored, yielding biosensors with limit of detections of 0.74 and 0.23 nm, and apparent affinity constants (KDGFET) of 6.7 and 8.8 nm, respectively. Both biosensing platforms show good specificity, fast time response, and wide dynamic range (0.1–100 nm). Moreover, SARS‐CoV‐2 spike protein is also detected in spiked nasopharyngeal swab samples. To rigorously validate this biosensing technology, the GFET response is matched with surface plasmon resonance measurements, exhibiting linear correlations (from 2 to 100 ng cm−2) and good agreement in terms of KD values. Finally, the performance of the biosensors fabricated through the nanoscaffold strategy is compared with those obtained through the widely employed monopyrene approach, showing enhanced sensitivity. The biofunctionalization of graphene field‐effect transistors through vinyl‐sulfonated polyethyleneimine nanoscaffold allows the immobilization of antifouling and recognition elements, yielding biosensors able to detect human ferritin and severe acute respiratory‐related coronavirus 2 spike protein in spiked nasopharyngeal swab samples. The developed technology is validated through the comparison with surface plasmon resonance measurements, and also with the widely employed monopyrene‐based biofunctionalization approach.