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

9725094 Murhammer爱荷华州和怀俄明州的主要研究者（PI）之间的这些合作提案的总体目标是通过在重组蛋白上创建更多的哺乳动物糖基化模式来开发昆虫细胞培养作为哺乳动物细胞培养的替代方案。 这将在该提案的三个主要目标中实现：草地贪夜蛾Sf-9细胞将被半乳糖基转移酶和唾液酸转移酶基因改变，并确定对模型蛋白SEAP（分泌型碱性磷酸酶）的影响。 将确定生物反应器环境对SEAP糖基化的影响，特别是二氧化碳和溶解氧浓度。 将对更复杂的蛋白质模型C1 INH 3进行类似的研究，C1 INH 3通常含有6个N-连接和7个O-连接的糖基化位点。 ***

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. ***