Characterization of N-Glycosylation Status of Micro-level Glycoprotein Using Solid Phase Extraction Based Techniques
A new strategy integrating the latest hydrophilic interaction chromatographic(HILIC) method with mass spectrometry technologies was developed for the enrichment and determination of intact glycopeptides and glycan structures from a glycoprotein.The average molecular weight,the ratio of glycosylation,the glycosite and the structures of glycoforms of ribonuclease B(RNaseB) were determined by MALDI-TOF-MS and ESI-LTQ-FT-MS.RNaseB contains 8.43% glycan and has a single glycosylation site at Asn34 with the glycan structure varied from five to nine mannose residues(Man5-9GlcNAc2).The strategy paves the way for systematic and confident analysis of complex glycoproteins with important biological or pharmaceutical functions.