Mass spectrometry-based glycoprofiling of biopharmaceuticals by using an automated data processing tool - SimGlycan®.

The therapeutic and immunological properties of biopharmaceuticals are governed by the glycoforms contained in them. Thus, bioinformatics tools capable of performing comprehensive characterization of glycans are significantly important to the biopharma industry. The primary structural elucidation of glycans using MS is tricky and tedious in terms of spectral interpretation. In this study, the biosimilars of a therapeutic monoclonal antibody and an Fc-fusion protein with moderate and heavy glycosylation, respectively, were employed as representative biopharmaceuticals for released glycan analysis using LC-MS/MS instead of conventional MS-based analysis. SimGlycan® - a software with proven ability to process tandem MS data for released glycans could identify eight additional glycoforms in Fc-fusion protein biosimilar, which were not detected during MS analysis of released glycans or glyco-peptide mapping of the same molecule. Thus, LC-MS/MS analysis of released glycans not only complements conventional LC-MS-based glycan profiling but can also identify additional glycan structures that may otherwise be omitted during conventional LC-MS based analysis of mAbs. The MS data processing tools, such as PMI Byos™ and SimGlycan® etc, can display pivotal analytical capabilities in automated LC-MS and LC-MS/MS-based glycan analysis workflows, especially for high-throughput structural characterization of glycoforms in biopharmaceuticals. This article is protected by copyright. All rights reserved.