Avidin–Biotin Technology in Gold Nanoparticle-Decorated Graphene Field Effect Transistors for Detection of Biotinylated Macromolecules with Ultrahigh Sensitivity and Specificity

The strong and specific noncovalent interaction between avidin and biotin is widely exploited in different types of enzyme-linked immunosorbent assay kits, labeled immunosensors, and polymer-based sensing devices for the detection of different biomarkers specific to different diseases such as cancer and influenza. Here, we employed the avidin–biotin technology in a novel gold nanoparticle-decorated graphene field-effect transistor (AuNP-GFET) and demonstrated the specific detection of the biotinylated macromolecules such as biotinylated proteins and nucleotides in the sub-picomolar (pM) range. The AuNP-GFET was constructed by fabricating six pairs of interdigital electrodes on graphene transferred on a SiO₂/Si substrate. The sensing performance of AuNP-GFET was characterized by the real-time two-terminal electrical current measurement upon injection of the analyte solution into a silicone pool preattached onto the electrodes. Avidin, a tetrameric biotin-binding protein with strong affinity and specificity, immobilized on AuNP-decorated single-layer graphene, was used as the sensing platform and transduced the electrical signal upon binding to the analyte macromolecules. The sensing capability of the AuNP-GFET was tested with the biotinylated protein A. Sensitivity of the present biosensor was estimated to be ∼0.4 pM. The specificity and applicability of the biosensor were confirmed using both synthetic and real samples. Because the biotin label can retain its binding capability to avidin with strong affinity and specificity even after conjugating with varieties of proteins and nucleotides, the present AuNP-GFET biosensor is expected to promote the research in developing different biosensors.