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Science 8 September 2006:
Vol. 313. no. 5792, pp. 1441 - 1443
DOI: 10.1126/science.1130256
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Reports

Humanization of Yeast to Produce Complex Terminally Sialylated Glycoproteins

Stephen R. Hamilton,1 Robert C. Davidson,1 Natarajan Sethuraman,1 Juergen H. Nett,1 Youwei Jiang,1 Sandra Rios,1 Piotr Bobrowicz,1 Terrance A. Stadheim,1 Huijuan Li,1 Byung-Kwon Choi,1 Daniel Hopkins,1 Harry Wischnewski,1 Jessica Roser,1 Teresa Mitchell,1 Rendall R. Strawbridge,2 Jack Hoopes,2 Stefan Wildt,1 Tillman U. Gerngross1,3*

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.

1 GlycoFi Inc., 21 Lafayette Street, Suite 200, Lebanon, NH 03766, USA.
2 Department of Surgery, Dartmouth-Hitchcock Medical Center, Lebanon, NH 03766, USA.
3 Thayer School of Engineering, Department of Biological Sciences and Department of Chemistry, Dartmouth College, 8000 Cummings Hall, Hanover, NH 03755, USA.

* To whom correspondence should be addressed. E-mail: tillman.gerngross{at}dartmouth.edu

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THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
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G.-C. Gil, W. H Velander, and K. E Van Cott (2008)
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Engineering of mucin-type human glycoproteins in yeast cells.
K. Amano, Y. Chiba, Y. Kasahara, Y. Kato, M. K. Kaneko, A. Kuno, H. Ito, K. Kobayashi, J. Hirabayashi, Y. Jigami, et al. (2008)
PNAS 105, 3232-3237
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N-Glycan Modification in Aspergillus Species.
E. Kainz, A. Gallmetzer, C. Hatzl, J. H. Nett, H. Li, T. Schinko, R. Pachlinger, H. Berger, Y. Reyes-Dominguez, A. Bernreiter, et al. (2008)
Appl. Envir. Microbiol. 74, 1076-1086
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Science. ISSN 0036-8075 (print), 1095-9203 (online)