Polyelectrolyte complexes as a tool for purification of plasmid DNA. Background and development |
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Authors: | Wahlund Per-Olof Gustavsson Per-Erik Izumrudov Vladimir A Larsson Per-Olof Galaev Igor Yu |
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Institution: | Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, PO Box 124, S-221 00 Lund, Sweden. |
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Abstract: | The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Plasmid DNA was selectively precipitated from a clarified alkaline lysate using the polycation poly(N,N'-dimethyldiallylammonium) chloride which formed insoluble polyelectrolyte complex (PEC) with the plasmid DNA. Soluble PECs of DNA with polycations have earlier been used for cell transformation, but now the focus has been on insoluble PECs. Both DNA and RNA form stable PECs with synthetic polycations. However, it was possible to find a range of salt concentration where plasmid DNA was quantitatively precipitated whereas RNA remained in solution. The precipitated plasmid DNA was resolubilised at high salt concentration and the polycation was removed by gel-filtration. |
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Keywords: | Purification Plasmid DNA Polyelectrolyte complexes |
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