Non-GMP plasmid production for transient transfection in bioreactors |
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Authors: | Georg Schmid Ernst-Jürgen Schlaeger Beat Wipf |
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Institution: | (1) Department of Biotechnology, F. Hoffmann-La Roche AG, Pharmaceutical Research – PRBT, Building 66/112A, CH-4070 Basel;(2) Department of Biotechnology, F. Hoffmann-La Roche AG, Pharmaceutical Research – PRBT, Building 66/112A, CH-4070 Basel |
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Abstract: | We describe a generic plasmid purification process for producing DNA for larger-scale transient transfection. Data on plasmid
quality with regard to residual protein, endotoxin content and presence of different plasmid forms is given. The effects of
contaminants and plasmid forms on expression levels of TNFRp55 and SEAP are discussed. Transient transfection of serum-free
suspension grown mammalian cells represents a suitable approach to provide research quantities of proteins (50–100 mg) within1–2
weeks.
This revised version was published online in July 2006 with corrections to the Cover Date. |
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Keywords: | endotoxin HEK293 plasmid purification SEAP transient transfection |
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