Abstract
Samples of 5 to 20 microg of human IgG were subjected to dithiothreitol treatment to reduce disulphide bridges, followed by tryptic digestion. Glycans released from the tryptic peptide mixture by PNGase F digestion were then derivatised with 2-aminoacridone. Labelled oligosaccharides were separated by normal-phase high-performance liquid chromatography and individual components were collected for matrix-assisted laser desorption ionization time-of-flight and electrospray mass spectrometric analysis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amidohydrolases / metabolism
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Aminoacridines
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Chromatography, High Pressure Liquid / methods*
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Disulfides / metabolism
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Dithiothreitol / pharmacology
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Glycoproteins / metabolism*
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Humans
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Immunoglobulin G / chemistry
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Immunoglobulin G / metabolism*
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Microchemistry
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Oligosaccharides / analysis
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Oligosaccharides / isolation & purification
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Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
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Polysaccharides / chemistry
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Polysaccharides / isolation & purification*
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Polysaccharides / metabolism
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
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Trypsin / metabolism
Substances
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Aminoacridines
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Disulfides
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Glycoproteins
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Immunoglobulin G
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Oligosaccharides
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Polysaccharides
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2-aminoacridone
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Trypsin
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Amidohydrolases
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Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
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Dithiothreitol