Effect of temperature on recombinant protein expression in Semliki Forest virus infected mammalian cell lines growing in serum- free suspension cultures |
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Authors: | Ernst-Jürgen Schlaeger Kenneth Lundstrom |
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Institution: | (1) Research Laboratories, F. Hoffmann-La Roche Ltd., CH-4070 Basel, Switzerland. |
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Abstract: | The firefly luciferase gene was introduced into the Semliki Forest virus (SFV) vector and high titer recombinant SFV particles
generated. The broad host range of SFV allowed efficient infection and high level expression of four mammalian cell lines
growing in serum-free suspension cultures. The incubation temperature had dramatic effects on the level and duration of recombinant
protein expression. For example, the luciferase activity was significantly higher in the rodent BHK and CHO cell lines incubated
at 33 °C compared to 37 °C when harvested 19 h post-infection. At 33 °C the specific expression levels increased 10–20 fold
during prolongation of the post-infection time up to 50 h. In contrast, a significant decrease in luciferase activity was
observed from 26 h post-infection for cell cultures incubated at 37 °C. Only a slight temperature effect on luciferase expression
was seen in the human cell line HEK293 and no effect was observed for the subclone293(EBNA).
This revised version was published online in August 2006 with corrections to the Cover Date. |
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Keywords: | expression levels low temperature Semliki Forest virus serum-free |
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