Glucan particles (GPs) are spherical hollow particles derived from Saccharomyces cerevisiae cell walls and mainly consist of β-1, 3-D-glucans. The inner hollow cavity of glucan particles can be loaded with different compounds, including protein antigens, and delivered to macrophages and dendritic cells. Moreover, the GP delivery system possesses β-glucan's intrinsic immunostimulatory properties. Therefore, GPs serve as both an antigen-presenting cell-targeted delivery system and an adjuvant.Here, we describe the production of GPs from S. cerevisiae using hot alkaline and solvent extraction and characterization of these particles for morphology, particle density, and hydrodynamic volume. A detailed protocol for loading and entrapping a model antigen, ovalbumin (OVA), into these particles using yeast RNA is presented. Similar methods are used to load pathogen-specific antigens (peptides, proteins, soluble extracts) which then can be tested in in vivo vaccination models.
Keywords: Cryptococcus; Dectin-1; Glucan particles; Vaccine production.