Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Production of Complex Human Glycoproteins in Yeast
Stephen R. Hamilton,*Piotr Bobrowicz,*Beata Bobrowicz,*Robert C. Davidson,*Huijuan Li,*Teresa Mitchell,*Juergen H. Nett,*Sebastian Rausch,*Terrance A. Stadheim,*Harry Wischnewski,*Stefan Wildt,*Tillman U. Gerngross
We report the humanization of the glycosylation pathway in theyeast Pichia pastoris to secrete a human glycoprotein with uniformcomplex N-glycosylation. The process involved eliminating endogenousyeast glycosylation pathways, while properly localizing fiveactive eukaryotic proteins, including mannosidases I and II,N-acetylglucosaminyl transferases I and II, and uridine 5'-diphosphate(UDP)-N-acetylglucosamine transporter. Targeted localizationof the enzymes enabled the generation of a synthetic in vivoglycosylation pathway, which produced the complex human N-glycanN-acetylglucosamine2-mannose3-N-acetylglucosamine2 (GlcNAc2Man3GlcNAc2).The ability to generate human glycoproteins with homogeneousN-glycan structures in a fungal host is a step toward producingtherapeutic glycoproteins and could become a tool for elucidatingthe structure-function relation of glycoproteins.
Thayer School of Engineering and the Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.
* Present address: GlycoFi, Inc., 21 Lafayette Street, Suite 200,Lebanon, NH 03766, USA.
To whom correspondence should be addressed. E-mail: tillman.gerngross{at}dartmouth.edu
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
|Abstract »|Full Text »|PDF »
The evolution of N-glycan-dependent endoplasmic reticulum quality control factors for glycoprotein folding and degradation.
S. Banerjee, P. Vishwanath, J. Cui, D. J. Kelleher, R. Gilmore, P. W. Robbins, and J. Samuelson (2007)
PNAS
104, 11676-11681
|Abstract »|Full Text »|PDF »
Fucosylation in prokaryotes and eukaryotes.
B. Ma, J. L. Simala-Grant, and D. E. Taylor (2006)
Glycobiology
16, 158R-184R
|Abstract »|Full Text »|PDF »
Humanization of yeast to produce complex terminally sialylated glycoproteins..
S. R. Hamilton, R. C. Davidson, N. Sethuraman, J. H. Nett, Y. Jiang, S. Rios, P. Bobrowicz, T. A. Stadheim, H. Li, B.-K. Choi, et al. (2006)
Science
313, 1441-1443
|Abstract »|Full Text »|PDF »
Fucose Removal from Complex-Type Oligosaccharide Enhances the Antibody-Dependent Cellular Cytotoxicity of Single-Gene-Encoded Bispecific Antibody Comprising of Two Single-Chain Antibodies Linked to the Antibody Constant Region.
A. Natsume, M. Wakitani, N. Yamane-Ohnuki, E. Shoji-Hosaka, R. Niwa, K. Uchida, M. Satoh, and K. Shitara (2006)
J. Biochem.
140, 359-368
|Abstract »|Full Text »|PDF »
A Model Vaccine Exploiting Fungal Mannosylation to Increase Antigen Immunogenicity.
J. S. Lam, M. K. Mansour, C. A. Specht, and S. M. Levitz (2005)
J. Immunol.
175, 7496-7503
|Abstract »|Full Text »|PDF »
In Vivo Synthesis of Mammalian-Like, Hybrid-Type N-Glycans in Pichia pastoris.
W. Vervecken, V. Kaigorodov, N. Callewaert, S. Geysens, K. De Vusser, and R. Contreras (2004)
Appl. Envir. Microbiol.
70, 2639-2646
|Abstract »|Full Text »|PDF »
To whom correspondence should be addressed. E-mail: 
