Comparison of denaturing agent effects in enzymatic N-glycan release for human plasma N-glycan analysis

Glycosylation is an essential posttranslational modification observed in the living proteome. profiles glycoproteins can change commonly diseases such as cancer. Identifying these changes crucial discovering new biomarkers for early diagnosis of One main steps N-glycan analysis to release N-glycans from by specific enzymes. The study compares common denaturing agent combinations used methods. In study, human plasma was test methods containing different detergent combinations. released were labeled with procainamide tag, purified using cellulose-containing solid-phase extraction cartridges, and analyzed high-performance liquid chromatography-hydrophilic interaction chromatography equipped fluorescence detection (HPLC-HILIC-FLD). results showed that sodium dodecyl sulfate deoxycholate (SDS + SDC) combination provided highest average FLD signal areas intensities analysis. protocol SDS resulted more reproducible intensities. It also found detected reduced when SDC 1,4-dithiothreitol (DTT) reducing agents. addition, deglycosylation various agents relatively minor variation N-glycosylation profiles.