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.