Structural Analysis of an Antigen Chemically Coupled on Virus‐Like Particles in Vaccine Formulation

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 formulations and their heterogeneous composition. Here we show that magic-angle spinning (MAS) solid-state NMR can be used to assess structure influenza virus hemagglutinin stalk long alpha helix antigen, both its free, unformulated form once chemically coupled surface large virus-like particles (VLPs). The sensitivity boost provided high-field dynamic nuclear polarization (DNP) proton detection at fast MAS rates allows overcome penalty associated with dilution. Comparison fingerprints between free VLP-coupled forms provides structural evidence conservation native fold upon bioconjugation. This work demonstrates high-sensitivity ripe play a major role design, formulation studies, manufacturing process development.