| Abstract [eng] |
Virus-like particles (VLPs) are nanostructures composed of self-assembling viral proteins. They mimic the structure of native viruses but are non-infectious, as they lack genetic material. Due to their safety and a wide range of functionalization methods, VLPs are promising tool for the delivery of various molecules. For VLP synthesis in this study, we selected the L-BC-1 virus found in Saccharomyces cerevisiae that lacks an extracellular phase. Previous research has focused on the structural characteristics and potential functions of yeast viruses within host cells. In this work, the stability and encapsulation potential of VLPs derived from the L-BC-1 virus was evaluated to assess their suitability for the development of VLP-based delivery systems. The recombinant capsid protein Gag of the L-BC-1 virus was successfully synthesized in Escherichia coli and S. cerevisiae, forming symmetrical and spherical particles approximately 40 nm in diameter. The stability of the Gag-derived VLPs was assessed using dynamic light scattering (DLS), transmission electron microscopy (TEM), and thermal shift assays (TSA). The particles maintained a stable size for up to 24 weeks under various conditions. Structural particle degradation was observed at high temperatures, in alkaline environment or upon exposure to urea. VLPs produced in E. coli exhibited greater sensitivity to environmental conditions compared to those synthesized in yeast. Using passive difussion, a small peptide nisin Z was encapsulated into the VLPs – resulting particles demonstrated antibacterial activity. Additionally, a large amount of red fluorescent mCherry protein was encapsulated via genetic fusion, and the ability of such particles to enter mammalian cells was demonstrated. It was shown that VLPs synthesized in bacteria exhibited cytotoxic effects on mammalian cells, while those produced in yeast did not. These findings provide valuable insights into the properties of the L-BC-1 virus and support its potential application in the development of delivery systems. |
