N-linked glycoprotein analysis using dual-extraction ultrahigh-performance liquid chromatography and electrospray tandem mass spectrometry.
Although reverse-phase liquid chromatography (RP-LC) is a common technique for peptide separation in shotgun proteomics and glycoproteomics, it often provides unsatisfactory results for the analysis of glycopeptides and glycans. This bias against glycopeptides makes it difficult to study glycoproteins. By coupling mass spectrometry (MS) with a combination of RP-LC and normal-phase (NP)-LC as an integrated front-end separation system, we demonstrate that effective identification and characterization of both peptides and glycopeptides mixtures, and their constituent glycan structures, can be achieved from a single sample injection event.
Siu
S O
SO
Department of Chemistry, The University of Hong Kong, Hong Kong, China.
Lam
Maggie P Y
MP
Lau
Edward
E
Yeung
William S B
WS
Cox
David M
DM
Chu
Ivan K
IK
D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D011134:Polysaccharides / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods; D012260:Ribonucleases / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014357:Trypsin / Q000737:chemistry
D006020:Glycopeptides; D006023:Glycoproteins; D011134:Polysaccharides; D020543:Proteome; D012709:Serum Albumin; D012997:Solvents; D006735:Horseradish Peroxidase; D012260:Ribonucleases; C029600:ribonuclease B; D014357:Trypsin