Large-Scale Production of Membrane Proteins in Pichia pastoris: The Production of G Protein-Coupled Receptors as a Case Study

  • Shweta Singh
  • Adrien Gras
  • Cedric Fiez-Vandal
  • Magdalena Martinez
  • Renaud Wagner
  • Bernadette Byrne
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 866)

Abstract

One of the major advantages of using Pichia pastoris is that it is readily adapted to large-scale culture in bioreactors. Bioreactors allow precise regulation of cell growth parameters increasing both yields and reproducibility of the culture. P. pastoris cultures grow to very high cell densities which helps minimise culture volume and facilitates downstream processing of the sample. Here, we provide protocols for the large-scale production of the human adenosine A2A receptor (A2AR) and provide some details of how bioreactor cultures can be used for optimisation of expression of the human dopamine D2 receptor (D2DR).

Key words

Large-scale expression Bioreactor Control of culture parameters Methanol probe Large-scale membrane preparations Receptor 

Notes

Acknowledgments

This research was funded by the MepNet consortium, the Biotechnology and Biological Research Council and GlaxoSmithKline.

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Copyright information

© Springer Science+business Media, LLC 2012

Authors and Affiliations

  • Shweta Singh
    • 1
  • Adrien Gras
    • 1
  • Cedric Fiez-Vandal
    • 2
  • Magdalena Martinez
    • 1
  • Renaud Wagner
    • 2
  • Bernadette Byrne
    • 1
  1. 1.Division of Molecular BiosciencesImperial College LondonLondonUK
  2. 2.Département Récepteurs et Protéines MembranairesCentre National de la Recherche Scientifique, Ecole Supérieure de Biotechnologie de StrasbourgIllkirchFrance

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