Fed-batch fermentation of GM-CSF-producing glycoengineered \u3ci\u3ePichia pastoris\u3c/i\u3e under controlled specific growth rate

  • Jacobs, Pieter P
  • Inan, Mehmet
  • Festjens, Nele
  • Haustraete, Jurgen
  • Van Hecke, Annelies
  • Contreras, Roland
  • Meagher, Michael M
  • Callewaert, Nico
Publication date
January 2010
Publisher
DigitalCommons@University of Nebraska - Lincoln
Language
English

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-glyc...

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