Biotechnology Letters

, Volume 34, Issue 4, pp 619–626 | Cite as

Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells

  • Yashas Rajendra
  • Divor Kiseljak
  • Lucia Baldi
  • David L. Hacker
  • Florian M. Wurm
Original Research Paper

Abstract

For most cultivated mammalian cells, glutamine is an essential medium component. However, glutamine consumption results in the production of ammonia, a cytotoxic byproduct. Here we investigated the effect of glutamine reduction on recombinant protein production and ammonia accumulation in transiently transfected CHO and HEK-293E cells maintained under conditions of growth arrest. Maximum transient recombinant protein yields were observed in HEK-293E cultures without glutamine and in CHO cultures with 2 mM glutamine. The initial concentration of glutamine correlated with the level of ammonia accumulation in each culture. For both a stable CHO-derived cell line and a polyclonal population of recombinant CHO cells grown under conditions of mild hypothermia, the highest volumetric protein productivity was observed in cultures without glutamine. Here, the level of ammonia accumulation also corresponded to the initial glutamine concentration. Our data demonstrate that reduction of glutamine in the medium is an effective approach to improve protein production in both transiently and stably transfected mammalian cells when applying conditions that reduce or arrest the growth of these cells.

Keywords

Ammonia CHO-DG44 Glutamine HEK-293E Stable gene expression Transient gene expression 

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Yashas Rajendra
    • 1
  • Divor Kiseljak
    • 1
  • Lucia Baldi
    • 1
  • David L. Hacker
    • 1
  • Florian M. Wurm
    • 2
  1. 1.Laboratory for Cellular Biotechnology (LBTC)École Polytechnique Fédéral de Lausanne (EPFL)LausanneSwitzerland
  2. 2.EPFL SV-IBI-LBTCLausanneSwitzerland

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