Humanization of Yeast to Produce Complex Terminally Sialylated Glycoproteins

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Science  08 Sep 2006:
Vol. 313, Issue 5792, pp. 1441-1443
DOI: 10.1126/science.1130256

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Yeast is a widely used recombinant protein expression system. We expanded its utility by engineering the yeast Pichia pastoris to secrete human glycoproteins with fully complex terminally sialylated N-glycans. After the knockout of four genes to eliminate yeast-specific glycosylation, we introduced 14 heterologous genes, allowing us to replicate the sequential steps of human glycosylation. The reported cell lines produce complex glycoproteins with greater than 90% terminal sialylation. Finally, to demonstrate the utility of these yeast strains, functional recombinant erythropoietin was produced.

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