Lectin-based SP3 technology enables N-glycoproteomics analysis

We developed a glycosylation enrichment method based on lectin and single-pot, solid-phase-enhanced sample preparation (SP3) technology, termed lectin-based SP3 technology (LectinSP3). LectinSP3 immobilized lectin on the SP3 beads for N-glycopeptide enrichment. It could identify over 1,100 N-glycosylation sites and 600 N-glycoproteins from 10 μg of mouse testis peptides. Furthermore, using LectinSP3 method, we characterized the N-glycoproteome of 1,000 mouse oocytes in three replicates, and identified a total of 363 N-glycosylation sites from 215 N-glycoproteins.