Characterization and optimization of vesicle properties in bioPISA : from size distribution to post-assembly loading

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Belluati, Andrea and Bloch, Adrian and Koynov, Kaloian and Nieva, Mariana Müller and Bagherabadi, Mohadeseh and Andrieu-Brunsen, Annette and Kolmar, Harald and Bruns, Nico (2024) Characterization and optimization of vesicle properties in bioPISA : from size distribution to post-assembly loading. Advanced Biology. 2400483. ISSN 2701-0198 (https://doi.org/10.1002/adbi.202400483)

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Abstract

This study investigates the formation and properties of vesicles produced via biocatalytic Polymerization-Induced Self-Assembly (bioPISA) as artificial cells. Methods for achieving size uniformity, including gentle centrifugation and sucrose gradient centrifugation, are explored, and the effects of stirring speed on vesicle morphology is investigated. The internal structure of the vesicles, characterized by a polymer-rich matrix, is analyzed using fluorescence correlation spectroscopy (FCS). Additionally, the feasibility of loading macromolecules into pre-formed vesicles is demonstrated using electroporation, and a fluorescent protein as well as enzymes for a cascade reaction were sucesfully incorporated into the fully assembled polymersomes. These findings provide a foundation for developing enzyme-synthesized polymeric vesicles with controlled morphologies for various applications, e.g., in synthetic biology.

ORCID iDs

Belluati, Andrea, Bloch, Adrian, Koynov, Kaloian, Nieva, Mariana Müller, Bagherabadi, Mohadeseh, Andrieu-Brunsen, Annette, Kolmar, Harald and Bruns, Nico ORCID logoORCID: https://orcid.org/0000-0001-6199-9995;
CORE (COnnecting REpositories)