earticle

영어

Glycosylation is one of the important post-translational modification of proteins, which have a variety of functions in a wide range of biological processes such as inflammation and immunization. Especially N-glycoproteins contain glycans attached to asparagine (N) residues. Dried Blood Spot (DBS) have a number of advantages which are easy to collection, transportation, and storage at room temperature compared with previous conventional sampling methods that store the blood in the liquid state at low temperature (-80 ℃). In this study, we performed direct analysis of the N-glycoproteins through optimization of sample preparation from human serum without any depletion and enrichment processes. Denaturing and desalting condition brfore MS analysis were tested and optimized conditions were applied to DBS to identify N-glycoproteins. We first found total 54 glycopeptides from 23 major glycoproteins in DBS directly. Among them, 17 site-specific N-glycopeptides from 11 major glycoproteins quantified by LC-MS/MS with GPA system [1]. The quantified N-glycopeptides have a similar retention time and CV% for quantitation was less than 30%. We were able to quantify neutral fucosylated and sialylated non-fucosylated glycan structures in DBS. Through these experiments, we confirmed that N-glycoproteins and N-glycopeptides analysis can be performed in dried blood spot directly. Further it is expected that N-glycoprotein analysis using dried blood spot can be applied to disease sample.