Thermo Fisher Scientific has developed a new approach that combines high-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) analysis, acid hydrolysis, and enzymatic digestion of protein glycans to determine information about glycan identity and terminal carbohydrate linkage isomers from small amounts of protein—just 0.5–1.6 μg per injection. Application Note (AN) 1050: Evaluating Protein Glycosylation in Limited-Quantity Samples by HPAE-PAD demonstrates that a suite of HPAE-PAD analyses can be performed with sample quantities as low as 3–10 μg. This direct determination approach saves time and reagent costs.
Cancer research frequently involves studying changes in protein glycosylation to identify important biomarkers such as prostate-specific antigen (PSA). The AN 1050 methodology uses an ion chromatography system configured for HPAE-PAD to investigate human PSA N-linked glycosylation and to evaluate the potential presence of O-linked glycans with methods optimized to work with small quantities of glycoproteins.