Manufacturing large plasmid DNA - new approaches

点击翻译按钮获取中文摘要

There has been a resurgence of interest in plasmid DNA-based vaccines and therapeutics after a relatively quiet period. In spite of large volume of data generated over the past two decades on plasmid processing, there are still challenges that persist when scaling up the production and purification of plasmid DNA, especially if the now projected high demand materializes specially for vaccination. These range from low productivity of microbial fermentation to difficult downstream processing and lack of a platform process.The current study builds on the vast body of publications and expertise in the group at UCL to address a few of the bottlenecks in the manufacturability of plasmid DNA. Hypothesis: tightly formed supercoiled DNA can potentially overcome the challenges of low quantity and quality of large supercoiled plasmids produced at scale.The study focuses on the most recent research in our labs where we have been successful in demonstrating that we can create tight supercoiled DNA with reduced size. This has been achieved in 6.5 Kb plasmids and we will extend this to larger plasmids (beyond 20Kb) where more information can be packaged. We will then examine the challenges of production in the fermenter followed by recovery and initial separation steps.