Fed-batch fermentation of GM-CSF-producing glycoengineered Pichia pastoris under controlled specific growth rate

Authors

Pieter P. Jacobs, Unit for Molecular GlycobiologyFollow
Mehmet Inan, University of Nebraska-LincolnFollow
Nele Festjens, Unit for Molecular Glycobiology
Jurgen Haustraete, Ghent University
Annelies Van Hecke, Unit for Molecular Glycobiology
Roland Contreras, Unit for Molecular Glycobiology
Michael M. Meagher, University of Nebraska-LincolnFollow
Nico Callewaert, Unit for Molecular GlycobiologyFollow

Date of this Version

2010

Citation

Jacobs et al. Microbial Cell Factories 2010, 9:93; http://www.microbialcellfactories.com/content/9/1/93

Comments

Copyright 2010 Jacobs et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License

Abstract

Background: Yeast expression systems with altered N-glycosylation are now available to produce glycoproteins with homogenous, defined N-glycans. However, data on the behaviour of these strains in high cell density cultivation are scarce.

Results: Here, we report on cultivations under controlled specific growth rate of a GlycoSwitch-Man5 Pichia pastoris strain producing Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) at high levels (hundreds of milligrams per liter). We demonstrate that homogenous Man₅GlcNAc₂ N-glycosylation of the secreted proteins is achieved at all specific growth rates tested.

Conclusions: Together, these data illustrate that the GlycoSwitch-Man5 P. pastoris is a robust production strain for homogenously N-glycosylated proteins.