on 05-03-201801:59 PM - edited on 10-15-202111:49 AM by Closed Account
Therese Wohlschlager 1,2, Kai Scheffler 2,3, Ines C. Forstenlehner 1,2,4, Wolfgang Skala 1,2, Stefan Senn 1,2, Eugen Damoc 5, Johann Holzmann 2,4 & Christian G. Huber 1,2 Nature Communications volume 9, Article number: 1713 (2018) doi:10.1038/s41467-018-04061-7 Robust manufacturing processes resulting in consistent glycosylation are critical for the efficacy and safety of biopharmaceuticals. Information on glycosylation can be obtained by conventional bottom–up methods but is often limited to the glycan or glycopeptide level. Here, we apply high-resolution native mass spectrometry (MS) for the characterization of the therapeutic fusion protein Etanercept to unravel glycoform heterogeneity in conditions of hitherto unmatched mass spectral complexity. Higher spatial resolution at lower charge states, an inherent characteristic of native MS, represents a key component for the successful revelation of glycan heterogeneity. Combined with enzymatic dissection using a set of proteases and glycosidases, assignment of specific glycoforms is achieved by transferring information from subunit to whole protein level. The application of native mass spectrometric analysis of intact Etanercept as a fingerprinting tool for the assessment of batch-to-batch variability is exemplified and may be extended to demonstrate comparability after changes in the biologic manufacturing process.
https://www.nature.com/articles/s41467-018-04061-7 1 Department of Biosciences, Bioanalytical Research Labs, University of Salzburg,
Hellbrunner Strasse 34, 5020 Salzburg, Austria.
2 Christian Doppler Laboratory for Innovative Tools for Biosimilar Characterization, University of Salzburg, Hellbrunner Strasse 34, 5020 Salzburg, Austria.
3 Thermo Fisher Scientific GmbH, DornierstraÃŸe 4, 82110 Germering, Germany.
4 Technical Development Biosimilars, Global Drug Development, Novartis, Sandoz GmbH,
Biochemiestrasse 10, 6250 Kundl, Austria.
5 Thermo Fisher Scientific GmbH, Hanna-Kunath-Strasse 11, 28199 Bremen, Germany.