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

病毒样颗粒（vlp）是一种大分子结构，适合作为预防和治疗用途的疫苗。其固有的高免疫原性已转移到这些嵌合VLPs表面的异源抗原上。此外，在一些研究中已经认识到装载货物的VLPs的巨大潜力。这些包括含有靶向给药的核酸的VLPs，因为它们可以用于基因治疗。嵌合型和载货型VLPs的结构、分散性和相行为对其功效和制造过程都至关重要。异源表位的引入和在治疗或宿主细胞核酸的背景下的加工显著影响其可制造性和加工路线。纯化过程中经常需要的分解反应以及随后的组装或衣壳稳定性在这里起着特别重要的作用。关于这些过程和性质对插入表位的依赖性以及对核酸装载的依赖性的认识不足。为了获得这些知识，必须在第一个资助期开发测量装置、方法和模型，借助这些装置、方法和模型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来详细研究VLPs在核酸加载或释放过程中的结构、分散性和相行为。因此，理解（反）组装反应和衣壳稳定性作为液相条件的函数，以及核酸的存在、浓度和类型，必须通过实验和计算机研究，采用并行的、选择性的和敏感的方法。这导致了从蛋白质水平（衣壳蛋白）到完全核酸负载VLP的跨尺度问题。因此，如果分子技术（MD， QSAR等）与机械工程模型相结合，生产过程的工艺工程描述才能成功。

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.