Characterization and Optimization of Vesicle Properties in bioPISA: From Size Distribution to Post-Assembly Loading

This study investigates the formation and properties of vesicles produced via biocatalytic Polymerization-Induced Self-Assembly (bioPISA) as artificial cells. We explore methods for achieving size uniformity, including gentle centrifugation and sucrose gradient centrifugation, and examined the effects of stirring speed on vesicle morphology. The internal structure of the vesicles, characterized by a polymer-rich matrix, is analysed using fluorescence correlation spectroscopy (FCS). Additionally, we demonstrate the feasibility of loading macromolecules into pre-formed vesicles using electroporation, successfully incorporating a fluorescent protein. Our findings provide a foundation for further developing polymeric vesicles, with controlled morphologies for various applications in synthetic biology.