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On-Demand Vaccine Production via Dock-and-Display of Biotinylated Antigens on Bacterial Extracellular Vesicles.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2843, pp. 195-216. - Publication Year :
- 2024
-
Abstract
- Engineered outer membrane vesicles (OMVs) derived from Gram-negative bacteria are a promising vaccine technology for developing immunity against diverse pathogens. However, antigen display on OMVs can be challenging to control and highly variable due to bottlenecks in protein expression and localization to the bacterial host cell's outer membrane, especially for bulky and complex antigens. Here, we describe methods related to a universal vaccine technology called AvidVax (avidin-based vaccine antigen crosslinking) for rapid and simplified assembly of antigens on the exterior of OMVs during vaccine development. The AvidVax platform involves remodeling the OMV surface with multiple copies of a synthetic antigen-binding protein (SNAP), which is an engineered fusion protein comprised of an outer membrane scaffold protein linked to a biotin-binding protein. The resulting SNAPs enable efficient decoration of OMVs with a molecularly diverse array of biotinylated subunit antigens, including globular and membrane proteins, glycans and glycoconjugates, haptens, lipids, nucleic acids, and short peptides. We detail the key steps in the AvidVax vaccine production pipeline including preparation and isolation of SNAP-OMVs, biotinylation and enrichment of vaccine antigens, and formulation and characterization of antigen-loaded SNAP-OMVs.<br /> (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Subjects :
- Bacterial Vaccines immunology
Bacterial Outer Membrane Proteins immunology
Bacterial Outer Membrane Proteins metabolism
Bacterial Outer Membrane Proteins genetics
Vaccine Development
Bacterial Outer Membrane metabolism
Bacterial Outer Membrane immunology
Extracellular Vesicles immunology
Extracellular Vesicles metabolism
Biotinylation
Antigens, Bacterial immunology
Antigens, Bacterial genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 2843
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 39141302
- Full Text :
- https://doi.org/10.1007/978-1-0716-4055-5_13