Electrochemical immunosensing of Crohn’s disease biomarkers using diazonium salt-grafted crystalline nanocellulose/carbon nanotube-modified electrodes

The first dual immunosensor is reported for the determination of IL-12 and IL-23, two relevant biomarkers of Crohn’s disease (CD). The strategy relies on the selective capture of the targets by the respective antibodies which were covalently immobilized onto SPCEs modified with crystalline nanocellulose (CNC) and multi-walled carbon nanotubes (MWCNTs) followed by conjugation with a detector antibody labelled with poly-HRP-Strept and amperometric transduction using the H₂O₂/HQ system. The combination of CNC, a nanomaterial scarcely exploited in immunosensing, with MWCNTs enables the preparation of a novel dual immunosensor for the determination of CD biomarkers in clinical samples, including faeces. The developed bioplatform exhibits a high reproducibility and selectivity allowing the simultaneous determination of IL-12 and IL-23 using a smaller sample volume and in a much shorter assay time than those reported for each target with commercial ELISA kits. The dual immunosensor was successfully applied to the analysis of raw serum and faeces spiked with IL-12 and IL-23 at the expected levels in patients with severe CD.