Multifunctional cerium nanolabels in electrochemical immunosensing with improved robustness and performance: determination of TIM-1 in colorectal cancer scenarios as a case study.

A multifunctional cerium oxide nanoparticles (CeO2NPs)-based nanolabel is exploited to implement an electrochemical sandwich-type immunoplatform for the determination of T-cell immunoglobulin and mucin domain 1 (TIM-1) biomarker, a mucin-like class I membrane glycoprotein associated with cancer angiogenesis. The immunoplatform is constructed using screen-printed electrodes where capture antibody is immobilized through the chemistry of diazonium salts. CeO2NPs exhibit robust pseudo-peroxidase activity even at high substrate concentrations. They are covalently functionalized in a simple manner after carboxylation with a detector antibody (dAb), acting dually as a nanozyme and nanocarrier for sensing bioreceptors. This allows the development of immunoplatforms with improved robustness and performance (in terms of a moderate enhancement in sensitivity, a significant expansion in the linear range, and a reduction in the background current) compared with the immunoplatforms prepared using nanolabels also decorated with the natural enzyme (horseradish peroxidase, HRP) or the conventional enzymatic labeling involving the dAb and an HRP-secondary antibody. Under the optimized experimental conditions, the developed electrochemical immunoplatform allows the highly sensitive detection of the TIM-1 glycoprotein, with a detection limit of 9.9 pg mL-1 and a linear working range of 33-600 pg mL-1. This performance permits biomarker quantification within clinically relevant ranges. This innovative configuration enables the precise diagnosis and stratification of colorectal cancer patients by analyzing plasma samples without pretreatment beyond a sample dilution and allows establishment of the first cut-off values reported for this purpose.