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Enzyme degradation, high performance liquid chromatography and liquid secondary ion mass spectrometry in the analysis of glycoproteins
Author(s) -
Smith Kevin D.,
Harbin AnneMarie,
Carruthers Robert A.,
Lawson Alexander M.,
Hounsell Elizabeth F.
Publication year - 1990
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130040612
Subject(s) - chemistry , chromatography , mass spectrometry , high performance liquid chromatography , oligosaccharide , fetuin , elution , glycoprotein , acetonitrile , liquid chromatography–mass spectrometry , glycopeptide , biochemistry , antibiotics
Analysis of small amounts of glycoproteins by high performance liquid chromatography (HPLC) and liquid secondary ion mass spectrometry (LSIMS) together with enzyme digestion has been investigated using fetuin as a model. Preliminary data indicates that 71% of the expected peptides were detected by LSIMS analysis of 200 pmol total digest. HPLC profiles of peptides and glycopeptides were obtained from 2 nmol of digest using a reversed phase ( C 18 ) column eluted in a solvent system containing TFA, water and acetonitrile. This has provided glycopeptides for subsequent oligosaccharide analysis. Strategies are reviewed for the chromatographic characterization of oligosaccharides following their release from glycopeptides by chemical and enzymatic procedures.