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Delauzun, Vincent; Amigues, Beatrice; Gaubert, Anais; Leone, Philippe; Grange, Magali; Gauthier, Laurent; Roussel, Alain
Methods, 08/2020, Volume: 180Journal Article
Display omitted •Extracellular vesicles production and their use for downstream applications.•Cell-surface membrane protein interaction studies using extracellular vesicles.•Extracellular vesicles as a tool for membrane protein structure determination.•Extracellular vesicles use for immunization and antibody generation. Producing intact recombinant membrane proteins for structural studies is an inherently challenging task due to their requirement for a cell-lipid environment. Most of the procedures developed involve isolating the protein by solubilization with detergent and further reconstitutions into artificial membranes. These procedures are highly time consuming and suffer from further drawbacks, including low yields and high cost. We describe here an alternative method for rapidly obtaining recombinant cell-surface membrane proteins displayed on extracellular vesicles (EVs) derived from cells in culture. Interaction between these membrane proteins and ligands can be analyzed directly on EVs. Moreover, EVs can also be used for protein structure determination or immunization purposes.
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