Nanohybrid nanozyme based colourimetric immunosensor for porcine gelatin

Enzyme mimicking nanomaterials, nanozymes, have gained considerable interest in the scientific community because of their superior properties compared to natural enzymes, including their high stability at extreme conditions, cheaper availability, and ease of synthesis. Herein, we report novel colloidal gold nanoparticles - graphene nanoplatelets - chitosan (CS) with peroxidase mimicking properties used to carry out highly sensitive and selective immunoassay for porcine gelatin detection. The interaction between anti-gelatin antibody conjugated nanozyme with porcine gelatin proteins produced an ultrasensitive immunoassay response in the form of a colourimetric signal directly proportional to the porcine gelatin protein concentration. The nanozyme produced a colourimetric response in the presence of its substrate, 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide (H₂O₂), demonstrating its peroxidase mimicking properties. The results revealed that the nanozyme exhibited remarkable selectivity and sensitivity in the assay, detecting proteins at concentrations as low as 86.42 pg/mL. Additionally, the immunosensor demonstrated a broad linear detection range spanning from 200 pg/mL to 2 ng/mL.