What is it about?
Production of plasmid DNA is important in the context of gene therapy and DNA vaccination. We developed a process based on tandem precipitation and multimodal chromatography that yields pure supercoiled plasmid DNA. The process involves E. coli culture, cell harvesting and alkaline lysis followed by isopropanol, ammonium acetate and PEG-8000 precipitation. Finally, sc pDNA is isolated by multimodal chromatography using a stepwise NaCl elution method.
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Why is it important?
Non-viral gene therapy and DNA vaccination currently hold great potential for treatment and prevention of genetic and acquired diseases. The large-scale manufacturing of plasmid DNA (pDNA) vectors, and the downstream processing in particular, is one of the key aspects of process development required to move non-viral gene therapy to the clinic. We use multimodal chromatography with an NaCl gradient to purify supercoiled plasmid DNA from open circular isoforms and RNA.
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This page is a summary of: A process for supercoiled plasmid DNA purification based on multimodal chromatography, Separation and Purification Technology, March 2017, Elsevier, DOI: 10.1016/j.seppur.2017.03.042.
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