Analysis of Glycopeptide Glycoforms in Monoclonal Antibody Tryptic Digest using a UPLC HILIC Column

This application note presents a UPLC HILIC/TUV/MS method for the separation of glycopeptides that is complementary to HILIC/FLR separation of N-linked glycans released from the protein. With this method, information about glycan heterogeneity and site occupancy is preserved and the same tryptic digest used for peptide mapping can be used. Since it does not require glycan release and labeling, complexity of sample preparation is reduced. This method is useful in the development and quality control of new protein-based therapies.