Detection and quantification of glycosylated peptides
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsAvraham Z. Rosenberg, Biao Shen
Life of this patent
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Abstract
A method of purification and/or separation of glycopeptides and quantitation of same. The method includes contacting a sample comprising glycopeptides to a hydrophilic enrichment substrate under conditions that permit the glycopeptides to bind to the hydrophilic enrichment substrate. The glycopeptides are eluted from the hydrophilic enrichment substrate with an ammonium formate and acetonitrile (ACN) in water solution to create an enriched glycopeptide sample, which may be subjected to analysis to identify specific glycopeptides.
Claims15
A method of characterizing and/or quantifying at least one glycopeptide in a sample containing at least one antibody, said method comprising:
- a) contacting the sample containing at least one antibody with one or more proteases to generate one or more glycopeptides;
- b) contacting said glycopeptides to a hydrophilic enrichment substrate under conditions that permit the glycopeptides to bind to the hydrophilic enrichment substrate;
- c) washing the hydrophilic enrichment substrate to remove non-glycopeptide contaminants from the hydrophilic enrichment substrate;
- d) eluting the glycopeptides from the hydrophilic enrichment substrate with an ammonium formate and acetonitrile (ACN) in water solution to create an enriched glycopeptide sample;
- e) applying the enriched glycopeptide sample to a separation column;
- f) eluting the glycopeptides from the separation column, thereby separating glycopeptides in the sample into at least one fraction;
- and
- g) subjecting said at least one fraction to mass spectrometry analysis to characterize and/or quantify said glycopeptides in said at least one fraction.
Dependencies are inferred from the claim text.