Capacitive nanobiosensing of β-1,4-galactosyltransferase-V colorectal cancer biomarker

Abstract

β-1,4-GalT-V is an enzyme with glycosyltransferase activity that glycosylates proteins and synthesizes the lactosylceramide sphingolipid in the Golgi apparatus. Colorectal cancer (CRC) tumor cells produce these biomolecules in high concentrations concerning normal cells, releasing them into the blood serum. Hence, β-1,4-GalT-V glycoprotein has emerged as a promising CRC biomarker, and its detection opens opportunities for the diagnosis/prognosis of CRC. We report the first capacitive nanobiosensor for the detection of β-1,4-GalT-V based on disposable screen-printed carbon electrodes (SPCEs) nanostructured with gold nanorods (AuNRs) and the Prussian blue (PrB) redox-active compound. AuNRs increased the SPCE surface area for antibody immobilization by physical adsorption and promoted the deposition of PrB onto the SPCE surface. Antibody-glycoprotein molecular biorecognition event onto the SPCE/AuNRs/PrB surface perturbed the PrB redox density-of-states and hence redox capacitance (Cμ̅). We found that changes in the inverse of Cμ̅ correlated well with the increasing concentrations of β-1,4-GalT-V in the linear range from 50 to 400 fM with a sensitivity of 1.5 µF fM-1 cm-2 and a limit of detection of 20 fM. The nanobiosensor detected the β-1,4-GalT-V glycoprotein from raw human serum samples with high specificity, ultrasensitivity, and reagentless label-free electrochemical transduction. The nanobiosensor could be applied as a CRC diagnosis/prognosis tool in a decentralized setting, with minimal patient sample manipulation and rapid response.