Fig. 3.

Prolonged ultracentrifugation (PUC) enables simultaneous recovery of secretory and extracellular vesicle-enriched glycoproteins from human plasma. A, Comparison of fractions recovered from 5 mL total plasma after 18 h or 2 h of UC. Yellow pellet recovered after PUC (18 h) was significantly larger and more visible than the pellet recovered after UC (2 h). B, Proportion of unique N-glycoproteins, N-glycopeptides, and N-glycosylated sites obtained when using PUC-ERLIC or standard UC-ERLIC approaches (both with PNGase F treatment). Significant glycoprotein enrichment was observed after PUC-ERLIC. The glycosylation percentage was calculated using the number of unique glycosylated products divided by the sum of all unique glycosylated and unmodified peptides.