Polyelectrolyte complexes as a tool for purification of plasmid DNABackground and development
| Autor: | Igor Yu. Galaev, Per-Olof Larsson, Vladimir A. Izumrudov, Per-Erik Gustavsson, Per-Olof Wahlund |
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| Rok vydání: | 2004 |
| Předmět: |
Electrophoresis
Agar Gel Plasmid preparation Lysis Chromatography Chemistry Clinical Biochemistry RNA DNA Cell Biology General Medicine Chromatography Ion Exchange Biochemistry Molecular biology Polyelectrolyte Analytical Chemistry Electrolytes chemistry.chemical_compound Transformation (genetics) Plasmid Calcium chloride transformation Chromatography Gel Electrophoresis Polyacrylamide Gel Plasmids |
| Zdroj: | Journal of Chromatography B. 807:121-127 |
| ISSN: | 1570-0232 |
| DOI: | 10.1016/j.jchromb.2004.01.046 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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