Carbohydrate release from picomole quantities of glycoprotein and characterisation of glycans by high-performance liquid chromatography and mass spectrometry

Samples of 5 to 20 mu g of human IgG were subjected to dithiothreitol treat ment to reduce disulphide bridges, followed by tryptic digestion. Glycans r eleased from the tryptic peptide mixture by PNGase F digestion were then de rivatised with 2-aminoacridone. Labelled oligosaccharides were separated by normal-phase high-performance liquid chromatography and individual compone nts were collected for matrix-assisted laser desorption ionization time-of- flight and electrospray mass spectrometric analysis. (C) 1999 Elsevier Scie nce B.V. All rights reserved.