Genetic Modifications and Environmental Factors Influencing Glycoprotein Processing in Sf-9 Cells (Collaborative Research)

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9725094 Murhammer The overall goal of these collaborative proposals between the Principal Investigators (PIs) at Iowa and Wyoming is to develop insect cell culture as an alternative to mammalian cell culture by creating a more mammalian glycosylation pattern on recombinant proteins. This will be accomplished in the three main aims of the proposal: Spodoptera frugiperda Sf-9 cells will be altered with genes for galactosyl transferase and sialyl transferase and the effect on the model protein SEAP (secreted alkaline phosphatase) determined. The effect of bioreactor environment on the SEAP glycosylation will be determined, especially carbon dioxide and dissolved oxygen concentration. Similar studies will be conducted on a much more complex protein model, C1INH, 3 which normally contains six N-linked and seven O-linked glycosylation sites. ***