One-third of the human-secreted proteome undergoes glycosylation, a post-translational modification that attaches carbohydrates to proteins (glycoproteins). Emerging findings showed that glycosylation is a molecular high-complex language enough to barcode a protein (Glyco-ID) and drive protein stability, cellular trafficking, and cell-surface interactions. Disease-associated deregulation of glycosylation can be directly dependent on genetic defects but also associated with metabolism, aging, and cancer. However, little is known about how the Glyco-ID of pathogenic biomarkers or cellular checkpoints could be used as a diagnostic and/or therapeutic target. This proposal aims 1) to identify aberrant Glyco-ID of biomarkers in patient-centered metabolic and neurodegenerative diseases; and 2) to understand the Glyco-ID of key immune cell checkpoints and its role in tumor immune evasion, in CAR-T therapy perspective. In this context, we have developed new approaches to studying protein-carbohydrate interactions using biochemical and glycobiology assays coupled with automated capillary-based immunoassays.