Gold nanocubes embedded biocompatible hybrid hydrogels for electrochemical detection of H₂O₂

Smart electrochemical biosensors have emerged as a promising alternative analytical diagnostic tool in recent clinical practice. However, improvement in the biocompatibility and electrical conductivity of the biosensor matrix and the immobilization of various bioactive molecules such as enzymes still remain challenging. The present research reports the synthesis of a biocompatible hydrogel network and its integration with gold nanocubes (AuNCs) for developing a novel biosensor with improved functionality. The interpenetrating hydrogel network consist of biopolymers developed using graft co-polymerization of β-cyclodextrin (β-CD) and chitosan (CS). The novelty of this work is in integrating the CS-g-β-CD hydrogel network with conductive AuNCs for improving hydrogel conductivity, biosensor sensitivity and use of the material for a biocompatible sensor. The present protocol advances the state of the art for the utilization of biopolymeric hydrogels system in synergy with an enzymatic biosensing protocol for exclusively detecting hydrogen peroxide (H₂O₂). Immobilization of the mitochondrial protein, cytochrome c (cyt c) into the hydrogel nanocomposite matrix was performed via thiol cross-linking. This organic-inorganic hybrid nanocomposite hydrogel matrix exhibited high biocompatibility (RAW 264.7 and N2a cell lines), improved electrical conductivity to attain high sensitivity (1.2 mA mM^-1 cm^-2) and a low detection limit (15 × 10^-9 M) for H₂O₂.