Abstract
The heterogeneity of recombinant glycoproteins expressed by CHO cells may partly result from a proteolytic activity present in cell culture supernatants. This study investigated the presence of proteases in CHO cell culture supernatants and their effects on the quality of a recombinant secreted viral envelope glycoprotein used as a model.
The glycoprotein characteristics were assessed by isoelectric focusing and western blot. Some of the isoforms of the glycoprotein so observed were found to be modified forms in particular at the NH2-terminal domain of the glycoprotein. Moreover, incubation of the intact glycoprotein with parental (untransfected) CHO cell culture supernatants resulted in similarly modified isoforms. Preliminary trials with protease inhibitors suggested that a serine protease could be involved in the glycoprotein modifications.
The presence of proteases in CHO cell culture supernatants might thus influence the quality of recombinant secreted glycoprotein and has to be considered in the optimisation of the cell culture conditions in large scale process development.
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© 1997 Springer Science+Business Media Dordrecht
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Tans, C., De Coninck, S.W., Gonze, MM., Fabry, L. (1997). Evaluation of the Proteolytic Activity Present in Cho Cell Culture Supernatants. In: Carrondo, M.J.T., Griffiths, B., Moreira, J.L.P. (eds) Animal Cell Technology. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-5404-8_46
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DOI: https://doi.org/10.1007/978-94-011-5404-8_46
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-6273-2
Online ISBN: 978-94-011-5404-8
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