Abstract
Physico-chemical parameters of calcium-phosphate DNA-coprecipitate formation were studied in detail using a simple centrifugation assay. Concentrations of the main reactants as well as the temperature and the reaction time were optimized. All parameters were verified in transfection experiments using intracellular (ß-galactosidase) or secreted proteins (tissue plasminogen activator) as indicators of transfection efficiency and expression in CHO or 293 HEK cells. It was demonstrated that controlling the particle size of the emerging calcium-phosphate crystals is the most critical parameter for obtaining high transfection efficiencies and recombinant protein expression.
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© 1995 Springer Science+Business Media Dordrecht
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Jordan, M., Wurm, F.M. (1995). High Level Transient Expression in Mammalian Cells: Identification and Optimization of Physico-Chemical Parameters of the Calcium Phosphate Transfection Method. In: Beuvery, E.C., Griffiths, J.B., Zeijlemaker, W.P. (eds) Animal Cell Technology: Developments Towards the 21st Century. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0437-1_8
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DOI: https://doi.org/10.1007/978-94-011-0437-1_8
Publisher Name: Springer, Dordrecht
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