Glycan characterization has remained a challenging aspect of biotherapeutic characterization compared to techniques such as intact mass or peptide map analysis, which most labs consider routine today. The addition of the glycan UPLC/FLR/MS workflow and use of the experimentally derived Waters Glycan GU Library within the Glycan Application Solution with UNIFI have addressed the desire for compliant-ready, automated, high-confidence glycan structure assignments by enabling rapid acquisition, review, and communication of individual glycan profile results, and the larger sets of glycan analyses used for comparability studies.

Additional capabilities with the Biopharmaceutical Platform Solution with UNIFI, such as glycan/glycopeptide DDA MS/MS analysis and the ability to execute exoglycosidase arrays, certainly complement this new workflow, providing additional orthogonal results that enable the characterization of even the most complex biotherapeutic glycoproteins.

In this application example we examine N-linked glycans from the innovator and a candidate biosimilar version of etanercept (Enbrel).

Related paper:
N- and O-Glycosylation Analysis of Etanercept Using Liquid Chromatography and Quadrupole Time-of-Flight Mass Spectrometry Equipped with Electron-Transfer Dissociation Functionality