Figure 4
From: Large-scale intact glycopeptide identification by Mascot database search
Workflow for N-glycoproteome analyses of serum samples. Briefly, the serum samples from control and patients diagnosed with prostate cancer were digested with trypsin and desalted with ZIC-HILIC SPE. Next, the N-linked sialylated glycopeptides were enriched with TiO2 beads, followed by LC-MS analysis using a Q Exactive mass spectrometer applying stepped NCE for HCD fragmentation. The intact glycopeptide mass spectra were submitted to the Mascot search engine for identification and relative quantification with Mascot Distiller. The data was searched against a custom glycoprotein database prepared from 21 linear N-linked sialylated glycans and proteins (444) known to be glycosylated in serum (PeptideAtlas N-Glyco build 2010).
