Abstract
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 ( KGFETDKDGFET<math> <mrow><msubsup><mi>K</mi> <mi>D</mi> <mrow><mi>G</mi> <mi>F</mi> <mi>E</mi> <mi>T</mi></mrow> </msubsup> </mrow> </math> ) 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 K D 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.