1. Structural Analysis of an Antigen Chemically Coupled on Virus-Like Particles in Vaccine Formulation
- Author
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Kaspars Tars, Olivier Ouari, Anna Kirsteina, Guido Pintacuda, Gilles Casano, Andris Kazaks, Kristaps Jaudzems, Anne Lesage, Janis Bogans, and Tobias Schubeis
- Subjects
Bioconjugation ,biology ,010405 organic chemistry ,Chemistry ,Manufacturing process ,Hemagglutinin (influenza) ,General Chemistry ,010402 general chemistry ,01 natural sciences ,Catalysis ,Virus ,0104 chemical sciences ,Solid-state nuclear magnetic resonance ,Virus-like particle ,Antigen ,biology.protein ,Biophysics ,Vaccines, Virus-Like Particle ,Antigens, Viral ,Nuclear Magnetic Resonance, Biomolecular ,Alpha helix - Abstract
Structure determination of adjuvant-coupled antigens is essential for rational vaccine development but has so far been hampered by the relatively low antigen content in vaccine formulations and by their heterogeneous composition. Here we show that magic-angle spinning (MAS) solid-state NMR can be used to assess the structure of the influenza virus hemagglutinin stalk long alpha helix antigen, both in its free, unformulated form and once chemically coupled to the surface of large virus-like particles (VLPs). The sensitivity boost provided by high-field dynamic nuclear polarization (DNP) and proton detection at fast MAS rates allows to overcome the penalty associated with the antigen dilution. Comparison of the MAS NMR fingerprints between the free and VLP-coupled forms of the antigen provides structural evidence of the conservation of its native fold upon bioconjugation. This work demonstrates that high-sensitivity MAS NMR is ripe to play a major role in vaccine design, formulation studies, and manufacturing process development.
- Published
- 2020