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

爱荷华州和怀俄明州的首席研究员（pi）之间的这些合作建议的总体目标是通过在重组蛋白上创建更接近哺乳动物的糖基化模式，开发昆虫细胞培养作为哺乳动物细胞培养的替代品。这将在三个主要目标中完成：用半乳糖转移酶和唾液转移酶基因改变Spodoptera frugiperda Sf-9细胞，并确定对模型蛋白SEAP（分泌碱性磷酸酶）的影响。测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。类似的研究将在一个更复杂的蛋白质模型上进行，C1INH 3通常包含6个n -链和7个o -链糖基化位点。＊＊＊

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. ***