Structure, conformation and phase changes of virus-like particles (VLPs) under chemical and mechanical stress in presence of nucleic acids

核酸存在下化学和机械应力下病毒样颗粒（VLP）的结构、构象和相变

• 批准号: 315439601
• 负责人: Professor Dr. Jürgen Hubbuch
• 金额: --
• 依托单位: Lehrstuhl für Molekulare Aufbereitung von Bioprodukten (MAB)
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/315439601?language=en
• 关键词: Structure; conformation; phase; changes; virus

Virus-like particles (VLPs) are macromolecular structures that are suitable as vaccines for prophylactic and therapeutic applications. Their high inherent immunogenicity has been transferred to heterologous antigens presented on the surface of these chimeric VLPs. Furthermore, the great potential of cargo-loaded VLPs has been recognized in several studies. These include VLPs containing nucleic acids for targeted administration as they could be used in gene therapy. The structure, dispersity and phase behaviour of chimeric and cargo-loaded VLPs is of central importance both for their efficacy and for the manufacturing process. The introduction of heterologous epitopes and the processing in the context of therapeutic or host cell nucleic acids significantly influence their manufacturability and the process route. The disassembly reaction frequently required for purification and the subsequent assembly or capsid stability play a particularly important role here.There is insufficient knowledge about the dependence of these processes and properties on the inserted epitope as well as on the loading with nucleic acids. In order to generate this knowledge, measurement set-ups, methods and models had to be developed in the first funding period, with the help of which the influence of the inserted epitope could be observed and described. So far, no such systematic approach exists for the detailed investigation of the structure, dispersity and phase behaviour of VLPs in the context of nucleic acid loading or release.The understanding of the (dis)assembly reaction and the capsid stability as a function of liquid phase conditions and the presence, concentration and type of nucleic acids must therefore be investigated experimentally and in silico with parallelized but also selective and sensitive methods. This results in cross-scale questions ranging from the protein level (capsid proteins) to complete nucleic acid-loaded VLP. A process-engineering description of the production processes can therefore only succeed if molecular techniques (MD, QSAR, etc.) are coupled with mechanistic engineering models.

病毒样颗粒（VLP）是适合作为疫苗用于预防和治疗应用的大分子结构。它们的高固有免疫原性已经转移到这些嵌合VLP表面上呈递的异源抗原。此外，一些研究已经认识到载货VLPs的巨大潜力。这些包括含有用于靶向施用的核酸的VLP，因为它们可用于基因治疗。嵌合和负载货物的VLP的结构、分散性和相行为对于它们的功效和制造过程都是至关重要的。异源表位的引入和在治疗性或宿主细胞核酸的情况下的加工显著影响其可制造性和工艺路线。纯化所需的拆卸反应和随后的组装或衣壳稳定性在此起着特别重要的作用。关于这些过程和性质对插入的表位以及对核酸负载的依赖性的知识还不够。为了产生这种知识，必须在第一个资助期内开发测量装置、方法和模型，在其帮助下，可以观察和描述插入表位的影响。到目前为止，还没有这样的系统方法存在于详细研究的结构，分散性和相行为的VLP在核酸加载或释放的情况下。理解（解）装配反应和衣壳稳定性作为液相条件和存在的函数，因此，核酸的浓度和类型必须用平行的但也是选择性和灵敏的方法进行实验和计算机模拟研究。这导致了从蛋白质水平（衣壳蛋白）到完整的核酸加载VLP的跨尺度问题。因此，只有在分子技术（MD、QSAR等）与机械工程模型相结合。