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Cytotechnology

, Volume 38, Issue 1–3, pp 109–117 | Cite as

Process development for functional membrane receptor production in mammalian cells

  • Christel Fenge
  • Irma Jansson
  • Thomas Fröberg
  • Marie Jönsson
  • Elke Lüllau
  • Linda Sygowski
  • Craig Moore
  • Dean Snyder
  • Michael Wood
Article

Abstract

Two model G-protein coupled membrane receptors (GPCRs), aserotonin (5HT) and a metabotropic glutamate (mGlu) receptor, stablyexpressed in CHO cells were used to characterize cultureconditions for maximum receptor expression and functionalactivity in membrane preparations. Expression levels of the5HT receptor were affected by the growth phase of the cellculture. Maximum receptor density, as measured by ligandbinding per mg membrane protein, was observed when cells wereharvested in late exponential growth phase. Expression couldbe increased further by addition of 10 mM sodium butyrate andincubation at 31 °C for 24 hours prior to cellharvest. In contrast, functional activity as determined byagonist-stimulated GTPγS binding was independent of the growthrate. For both receptors, butyrate treatment at decreasedtemperature negatively affected functional activity. The mGlureceptor membranes lost functional activity considerably whenthe cells were cultured in an agitated system either onmicrocarriers or as aggregates in suspension. Functionalactivity could be restored and further improved compared to acontrol grown in T-flasks when the cell culture was incubatedat 31 °C for 48 hours following a complete mediumexchange and omission of sodium butyrate.

butyrate CHO G-protein coupled receptor low temperature and suspension culture 

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Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Christel Fenge
    • 1
  • Irma Jansson
    • 1
  • Thomas Fröberg
    • 1
  • Marie Jönsson
    • 1
  • Elke Lüllau
    • 1
  • Linda Sygowski
    • 2
  • Craig Moore
    • 2
  • Dean Snyder
    • 2
  • Michael Wood
    • 2
  1. 1.AstraZeneca Biotech LaboratorySödertäljeSweden
  2. 2.AstraZeneca PharmaceuticalsWilmingtonUSA

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