Manufacturing large plasmid DNA - new approaches

制造大质粒 DNA——新方法

• 批准号: 2065248
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2065248
• 关键词: Manufacturing; large; plasmid; DNA; new

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.

在相对平静的时期之后，对基于质粒DNA的疫苗和治疗剂的兴趣重新抬头。尽管在过去的二十年中产生了大量关于质粒加工的数据，但当扩大质粒DNA的生产和纯化时，仍然存在挑战，特别是如果现在预计的高需求专门用于疫苗接种的话。这些问题包括微生物发酵的低生产率、下游加工困难以及缺乏平台过程。目前的研究建立在UCL小组的大量出版物和专业知识的基础上，以解决质粒DNA可制造性的一些瓶颈。假设：紧密形成的超螺旋DNA可以潜在地克服大规模生产的大超螺旋质粒的低数量和质量的挑战。这项研究集中在我们实验室的最新研究中，我们已经成功地证明我们可以创造出尺寸减小的紧密超螺旋DNA。这已经在6.5 Kb质粒中实现，我们将把它扩展到更大的质粒（超过20 Kb），在那里可以包装更多的信息。然后，我们将研究在发酵罐中生产的挑战，然后是回收和初始分离步骤。