Method for identification of glycosylated polypeptides

G - Physics – 01 – N

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G01N 33/68 (2006.01)

Patent

CA 2527650

The instant invention provides a stepwise method for identification of glycosylated polypeptides. The steps of the method involve treatment of a biological sample containing a glycosylated polypeptide with at least one protease to begin peptide digestion, followed by deglycosylation and subsequent re-digestion with at least one protease to result in deglycosylated polypeptide fragments. The deglycosylated polypeptide fragments are then sequenced using mass spectrometry and identified by sequence comparison with a database of known sequences.

La présente invention concerne une méthode progressive d'identification de polypeptides glycosylés. Les étapes de cette méthode comprennent le traitement d'un échantillon biologique contenant un polypeptide glycosylé avec au moins une protéase pour commencer la digestion des peptides, puis la déglycosylation et la digestion subséquente avec au moins une protéase de manière à parvenir à des fragments de polypeptides déglycosylés. Ces fragments de polypeptides déglycosylés sont ensuite séquencés au moyen d'une spectrométrie de masse et identifiés par une comparaison de séquences avec une base de données de séquences connues.

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