Propagation ofSpodoptera frugiperdacells (Sf9) and production of recombinant proteins with the baculovirus expression system using improved spinner flasks with membrane aeration
✍ Scribed by Volker Jäger; Anja Kobold
- Publisher
- Springer-Verlag
- Year
- 1995
- Tongue
- English
- Weight
- 322 KB
- Volume
- 9
- Category
- Article
- ISSN
- 0951-208X
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✦ Synopsis
It has been shown that the growth of Spodoptera frugiperda cells is significantly reduced or ceased under oxygen limiting culture conditions. This paper describes the USC of a new membrane-aerated spinner flask which was compared to conventional surface-aerated spinner flasks with regard to growth of the insect cell line Sf9 and recombinant protein production after infection with baculovirus. Using a commercially available strum-free culture medium Sf9 cells reached highest cell densities (3x10" ml-') in the membrane-acrated spinner flask. Production of recombinant protein was also influenced by the oxygen supply. In the membrane-acrated spinner flask and in a surface-aerated spinner flask with reduced filling volume more than 20000 U ml-' of a recombinant interleukin-2 variant were accumulated whereas only 100 U ml-' were produced in a surface-aerated spinner flask with insufficient oxygen supply. Sufficient oxygenation appears to be essential for proliferation of St9 cells as well as recombinant protein production after infection with baculovirus. Membrane oxygenation allows sufficient oxygen supply at high cell density and an at least 2.5 fold higher filling volume per spinner unit.