Fractionation of ovalbumin glycopeptide AC-C by high-pressure liquid chromatography. Determination of structure by 1H-NMR spectroscopy.

We describe for the first time a method for the separation of intact ovalbumin glycopeptides by high-pressure liquid chromatography (HPLC). The separation was achieved using two reverse-phase columns connected in series and eluting with an isocratic solvent system at acid pH containing 1-hexane sulfonate. Ovalbumin glycopeptide fraction AC-C has been separated into at least four distinct glycopeptides. High resolution 1H-NMR spectroscopy has confirmed the reported structure of the two major species. We also extend our structural studies to the two other glycopeptides and establish the structure of a previously unreported ovalbumin glycopeptide, Man3GalGlcNAcAsn.