Abstract
Recent investigations have implicated aberrant glycosylations in various malignancies, including epithelial ovarian cancer (EOC). The protocol here identifies O-linked carbohydrate patterns in EOC plasma glycoproteins through chemical cleavage and purification of these glycans. Dialyzed plasma is subjected to reductive beta-elimination with alkaline borohydride to release O-linked oligosaccharides from glycoproteins. Enrichment of released glycans, as well as removal of peptide and other contaminants, is followed by carbohydrate pattern analysis with MALDI-FT-ICR-MS.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Blood Proteins / chemistry*
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Blood Proteins / metabolism
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Borohydrides / chemistry
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Dialysis / methods
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Female
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Glycosylation
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Hexosamines / analysis
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Hexoses / analysis
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Hexuronic Acids / analysis
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Humans
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Middle Aged
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Ovarian Neoplasms / blood*
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Polysaccharides / analysis*
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Polysaccharides / chemistry
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Polysaccharides / isolation & purification
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Pronase / metabolism
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Solid Phase Extraction / methods
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
Substances
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Blood Proteins
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Borohydrides
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Hexosamines
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Hexoses
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Hexuronic Acids
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Polysaccharides
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sodium borohydride
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Pronase