Abstract
The presence and structure of oligosaccharides in glycoproteins are known to influence the biological properties of therapeutic proteins. The ability to characterize the oligosaccharides in real-time during animal cell cultivation offers the possibility to control and ensure the quality and consistency of the therapeutic proteins. This paper presents a number of real-time analytical techniques in the quantitative measurement of macro- and microheterogeneity of protein glycosylation. The monitoring techniques include tandem microbore chromatography, capillary electrophoresis, continuous peptide digestion and MALDI/TOF mass spectrometry. The results from these real-time measurements during the cultivation of a recombinant Chinese Hamster Ovary cell culture encoding for human interferon-γ (IFN-γ) will be presented. Rapid and real-time analysis (4 hours) and high sensitivity (0.5 µg) have been achieved to characterize both macro- and microheterogeneity of IFN-γ glycosylation. Both the site occupancy and fractions of biantennary, triantennary and tetraantennary structures of inteferon-γ were found to remain unchanged during culture. However, sialylation percentage of IFN-γ remained relatively constant in the early stage of the culture, and dropped steadily after massive cell death. This decrease in product sialylation was later confirmed to result from extracellular sialidase activity.
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© 1998 Springer Science+Business Media Dordrecht
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Gu, X., Harmon, B.J., Wang, D.I.C. (1998). Monitoring and Characterization of Glycoprotein Quality in Animal Cell Cultures. In: Galindo, E., Ramírez, O.T. (eds) Advances in Bioprocess Engineering. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-0643-8_1
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DOI: https://doi.org/10.1007/978-94-017-0643-8_1
Publisher Name: Springer, Dordrecht
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