Impact of storage on the stability and the protective effect of extracellular vesicles released by <i>Candida albicans</i>

Extracellular vesicles (EVs) released by <i>Candida albicans</i> are promising vaccine prototypes due to their multi-antigenic nature. However, their storage and handling conditions are not well understood, raising concerns about their biotechnological use. This study evaluated the long-term physical and functional stability of <i>C. albicans</i> EVs. We compared three isolation methods to identify the most effective one and suggested a quality control measure based on protein and sterol ratios. We also examined the effects of vacuum drying and freeze-thaw cycles on EV stability. Our findings show that <i>C. albicans</i> EVs maintain their biological function after long-term storage at -80°C and under various conditions. Notably, their protective effect in an insect model was reduced through repeated freeze-thaw cycles. This research provides valuable insights for the efficient use of these vesicles in future studies.