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

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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.