As one of the most important posttranslational modifications, glycosylation plays critical roles in protein folding, trafficking, cell differentiation, immune recognition, etc. The alteration of glycosylation is closely associated with the pathological processes during and after caner development, and thus holds great value in cancer detection. In this chapter, we describe a protocol on the glycosylation profiling of tumor marker in plasma using bead-based immunoassay with CA125 as a model, including bead coupling, coupling control, glycosylation assay, as well as the plasma screening for breast cancer patients. This protocol can be used to profile the glycosylation of protein markers in clinical plasma or serum samples for different human cancers.
