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Humanization of Yeast to Produce Complex Terminally Sialylated Glycoproteins

Author:
Hamilton, Stephen R., Davidson, Robert C., Sethuraman, Natarajan, Nett, Juergen H., Jiang, Youwei, Rios, Sandra, Bobrowicz, Piotr, Stadheim, Terrance A., Li, Huijuan, Choi, Byung-Kwon, Hopkins, Daniel, Wischnewski, Harry, Roser, Jessica, Mitchell, Teresa, Strawbridge, Rendall R., Hoopes, Jack, Wildt, Stefan, Gerngross, Tillman U.
Source:
Science 2006 v.313 no.5792 pp. 1441-1443
ISSN:
0036-8075
Subject:
engineering, Komagataella pastoris, humans, glycosylation, recombinant proteins, yeasts, genes, erythropoietin
Abstract:
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.
Agid:
1531531