At-line high throughput site-specific glycan profiling using targeted mass spectrometry.

Autor:	Chi B; MedImmune, Milstein Building, Granta Park, Cambridge, CB21 6GH, UK., Veyssier C; MedImmune, Milstein Building, Granta Park, Cambridge, CB21 6GH, UK., Kasali T; MedImmune, Milstein Building, Granta Park, Cambridge, CB21 6GH, UK., Uddin F; MedImmune, Milstein Building, Granta Park, Cambridge, CB21 6GH, UK., Sellick CA; MedImmune, Milstein Building, Granta Park, Cambridge, CB21 6GH, UK.
Jazyk:	angličtina
Zdroj:	Biotechnology reports (Amsterdam, Netherlands) [Biotechnol Rep (Amst)] 2020 Jan 22; Vol. 25, pp. e00424. Date of Electronic Publication: 2020 Jan 22 (Print Publication: 2020).
DOI:	10.1016/j.btre.2020.e00424
Abstrakt:	Protein post-translational modification (PTM) plays an important role in many biological processes; of which glycosylation is arguably one of the most complex and diverse modifications and is crucial for the safety and efficacy of biotherapeutic proteins. Mass spectrometric characterization of protein glycosylation is well established with clear advantages and disadvantages; on one hand it is precise and information-rich, as well as being relative inexpensive in terms of the reagents and consumables despite the instrumentation cost and, depending on the method, can give site specific information; on the other hand it generally suffers from low throughput, restriction to largely purified samples and is less quantitative, especially for sialylated glycan species. Here, we describe a high throughput, site-specific, targeted mass spectrometric peptide mapping approach to quickly screen/rank candidate production cell lines and culture conditions that give favourable glycosylation profiles directly from conditioned culture media for an Fc-fusion protein. The methodology is fully compatible with automation and combines the speed of 'top-down' mass spectrometry with the site-specific information of 'bottom-up' mass spectrometry. In addition, this strategy can be used for multi-attribute product quality screening/monitoring as an integral part of cell line selection and process development. Competing Interests: The authors declare no financial or commercial conflict of interest. (© 2020 The Authors.)
Databáze:	MEDLINE
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