Magnetic nanoparticles coated with maltose-functionalized polyethyleneimine for highly efficient enrichment of N-glycopeptides.

Hydrophilic interaction chromatography (HILIC) adsorbents have drawn increasing attention in recent years due to their high efficiency in N-glycopeptides enrichment. The hydrophilicity and binding capacity of HILIC adsorbents are crucial to the enrichment efficiency and mass spectrometry (MS) detection sensitivity of N-glycopeptides. Herein, magnetic nanoparticles coated with maltose-functionalized polyethyleneimine (Fe3O4-PEI-Maltose MNPs) were prepared by one-pot solvothermal reaction coupled with "click chemistry" and utilized for N-glycopeptides enrichment. Owing to the presence of hydrophilic and branched polyethyleneimine, the amount of immobilized disaccharide units was improved about four times. The N-glycopeptides capturing capacity was about 150mg/g (IgG/MNPs) and the MS detection limitation as low as 0.5fmol for IgG and 85% average enrichment recovery were feasibly achieved by using this hybrid magnetic adsorbent. Finally, 1237 unique N-glycosylation sites and 1567 unique N-glycopeptides from 684 N-glycoproteins were reliably characterized from 60μg protein sample extracted from mouse liver. Therefore, this maltose-functionalized polyethyleneimine coated adsorbent can play a promising role in highly efficient N-glycopeptides enrichment for glycoproteomic analyses of complex protein samples.