Gene regulation in baculovirus-infected insect cells: application in the development of a budded virus-free expression system for the production of VL

基于vlp的疫苗具有保护人类（和动物）免受某些最严重病毒性疾病侵害的巨大潜力。杆状病毒表达系统以其产生高产量高质量VLPs的能力而闻名，这与易于使用和可扩展性相结合，导致许多医疗保健/生物技术公司探索将其用于疫苗开发。然而，事实证明具有挑战性的是去除污染的杆状病毒颗粒，这增加了下游加工和最终成功推出新疫苗所涉及的成本、复杂性和时间。该项目有可能对上游生产阶段进行改进，从而消除对复杂且耗时的下游加工的需求。潜在的影响将是简化和降低VLP纯化的成本，最终可能导致更有效的基于VLP的疫苗管道。该项目旨在建立在我们对杆状病毒颗粒（出芽病毒，BV）产生的基因调控的基本认识的基础上，这发生在病毒复制周期的后期阶段。目的是使表达载体系统的发展，其中BV合成可以在VLP生产过程中下调。因此，该项目包含一个基本的研究元素：了解杆状病毒复制周期后期的基因调控与BV的产生有关。但它也将包括一个工业元素，将利用这些信息开发一种新的表达系统，在生产过程中可以关闭或下调BV的生产，以产生用于纯化的VLPs。合作组织OET有限公司在开发和商业化新型杆状病毒表达系统方面拥有丰富的经验，但开发一种抑制细菌性病毒生产的系统需要对细菌性病毒生产的基因调控有更深入的了解，以便可以对其进行操纵并转化为商业上可行的表达载体。除了DTP培训外，学生还将接受现代生物科学技术的具体培训，包括DNA， RNA和蛋白质分析，克隆，生物信息学，光学和电子显微镜，病毒处理和细胞培养。学生将通过两周一次的实验室会议、期刊俱乐部、研讨会和科学会议上的演讲（海报和演讲）来发展他们的沟通和演讲技巧。学生也被鼓励参加示范，以发展教学技能，并通过布鲁克斯公共参与网络参加外展活动。此外，学生将有机会跟随OET的一位高级科学家进行合同研究，以了解合同研究在许多英国和海外制药和生物技术公司领导的疫苗和其他研发项目中的作用。这将有助于了解从早期研发阶段进入试验和生产阶段所面临的挑战。

VLP-based vaccines have significant potential to protect people (and animals) from some of the most serious viral diseases. The baculovirus expression system is known for its ability to produce high yields of quality VLPs and this combined with ease of use and scalability has led to many healthcare/biotechnology companies exploring their use in vaccine development. What has proved challenging, however, is the removal of contaminating baculovirus particles and this has added to the costs, complexities and time involved in downstream processing and ultimate successful launch of new vaccines. This project has potential to deliver improvements to the upstream production phase that will negate the need for complex and time consuming downstream processing. The potential impact will be to simplify and reduce the cost of VLP purification that may ultimately result in a more efficient VLP-based vaccine pipeline.This project aims to build on our basic understanding of the regulatory control of the genes involved in baculovirus particle (budded virus, BV) production, which occurs in the late phase of the virus replication cycle. The aim is to enable development of an expression vector system in which BV synthesis can be down regulated during VLP production. The project therefore contains a basic research element: understanding gene regulation in the late phase of the baculovirus replication cycle as pertinent to BV production. But it will also include an industrial element that will use the information to develop a novel expression system in which BV production can be switched off or down-regulated during production-runs to yield VLPs for purification. The collaborating organisation, OET Ltd, has a wealth of experience in developing and commercialising novel baculovirus expression systems but development of a system to knock down BV production requires greater understanding of the gene regulation of BV production so that this can be manipulated and translated into a commercially-viable expression vector. In addition to DTP training, the student will receive specific training in modern bioscience techniques including DNA, RNA and protein analyses, cloning, bioinformatics, light and electron microscopy, virus handling and cell culture. The student will be supported to develop their communication and presentation skills through fortnightly lab meetings, a journal club, seminars and presentations (poster and talks) at scientific meetings. Students are also encouraged to take part in demonstrating to develop teaching skills and to take part in outreach activities through the Brookes Public Engagement Network.In addition, the student will benefit from an opportunity to shadow one of OET's senior scientists conducting contract research to obtain an understanding of the role of contract research in vaccine and other R&D programmes led by many of the UK's and overseas' pharma and biotechnology companies. This will provide an insight into the challenges of moving from early stage R&D into trials and production.