Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells.
In the last decades, the number of approved therapeutic proteins drugs is increasing exponentially and a large number of new therapeutic entities are progressing through clinical trials, solidifying biologics as the most promising class of pharmaceuticals on the market. Several cell lines are available for biopharmaceutical processes but mammalian cells are preferred since they give fewer problems for immunogenicity as they produce human-like post-translational modifications (PTMs). Glycosylation is the most common and complex (for both bioprocess engineering and quality control) of these modifications. Obtaining the desired glycosylation pattern is crucial for therapeutic proteins as it can impact significantly stability, half-life and safety as well as driving molecular processes, modifying the way drug interacts with patients' cells. As a consequence, glycosylation (like other PTMs) needs to be regulated and accurately analyzed during biopharmaceutical production. Herein we describe and discuss the analytical approaches for glycosylation analysis of therapeutic glycoproteins produced in CHO (Chinese Hamster Ovary) cells. This chapter will describe glycoprotein purification after separation from producing cell lines, N-glycan release and their variants fine structural characterization through mass spectrometry techniques.
['Carillo|Sara|S|', 'Mittermayr|Stefan|S|', 'Farrell|Amy|A|', 'Albrecht|Simone|S|', 'Bones|Jonathan|J|']
D000818:Animals D001688:Biological Products D016466:CHO Cells D018929:Cell Culture Techniques D006224:Cricetinae D003412:Cricetulus D006031:Glycosylation D006801:Humans D013058:Mass Spectrometry D011134:Polysaccharides D015202:Protein Engineering D011499:Protein Processing, Post-Translational D011994:Recombinant Proteins
CHO cells Mass spectrometry Glycan analysis Liquid chromatography Monoclonal antibodies