Glycosylation can have significant effects on the clinical safety and efficacy of biopharmaceuticals [1,2]. As a result, both innovator drugs’ and biosimilars’ glycan profiles need to be fully characterized during the product lifecycle, from development to commercialization including batch-to-batch consistency and release to the market . Additionally, a vast amount of research has been performed on the identification of disease biomarkers based on the glycosylation of biological samples [3,4,5]. Due to the high demand and institutional pressure for biopharmaceutical characterization  and the huge number of samples that a clinical setting is exposed to , there is a need for the development of a rapid, sensitive and selective method for glycan and glycopeptide analysis. Glycosylation analysis both at the N-glycan and glycopeptide level provide complementary information. N-glycan analysis gives specific information about the glycan structure and linkages by exoglycosidases analysis while glycopeptide analysis is a more favourable approach for the characterization of site-specific glycan compositions.
A new label for both N-glycan and glycopeptide analysis has been investigated. This label has an imidazolium group with a permanent positive charge that has been used as MS probe showing greater spectral peak intensities and lower limits of detection . This new label synthesis is achieved in two steps. Glycoanalysis with this tag is quick and easy: it just requires 4 steps and no dry down steps are involved in the process, allowing it to be completed in as little as 2 hours. Final sample separation and identification is performed by tandem HILIC-UHPLC and ESI-MS.
This poster will focus on the details of this new chemical derivatisation label. We will show that reliable data for both N-glycan and glycopeptide identification is generated using this rapid procedure with a small amount of glycoprotein (less than 10 μg for proteins with only one glycosylation site). The results obtained as a proof-of-concept for the analysis of antibodies and fetuin will be presented. As further optimization is required, the plans for future work will also be discussed.
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