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AssigneeGlaxosmithkline Biologicals SA
InventorsOlivier LE BUSSY, Baptiste PIERMAN, Marc ORVAL, Delphine JASON
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This application discloses a method for purifying pDNA, particularly pDNA that that can be used to produce RNA, the RNA preferably encoding a therapeutic or immunogenic peptide or polypeptide. The pDNA can be grown in a bacteria such as E. coli by culturing or fermenting bacteria containing the plasmid and obtaining and purifying the pDNA. The present method allows the pDNA to be obtained in high yield and with high purity. In one embodiment of the invention, the level of all non-pDNA materials can be significantly reduced by the process. In some embodiments, the ratio of supercoiled plasmid DNA (scDNA) to non-supercoiled pDNA (non-scDNA, such as open circular plasmid DNA (ocDNA)) can be increased by one or more process steps that separate or allow for separation of scDNA and ocDNA or process steps that increase the amount of scDNA to ocDNA.
A method of purifying plasmid DNA (pDNA), comprising the steps of:
A method of improving the quality of a template pDNA prior to an in vitro transcription reaction, comprising the steps of:
A method for purifying pDNA comprising the steps of:
A plasmid DNA composition comprising pDNA wherein at least 80% of the plasmid DNA is in supercoiled form, less than 15% is in open-circular form and less than 5% is in other isoforms, all percentages being based on the total amount of pDNA present.
Dependencies are inferred from the claim text.