Alpha-BET: Functional labeling of envelope glycoproteins with single domain antibodies for in-virus single molecule imaging
Williamson, D. J.; Zaza, C.; Carlon Andres, I.; Gentili, A.; Daly, J. I.; Holmes, H.; Thrush, J.; Starling, T.; Neil, S.; Owens, R.; Malim, M. M.; Tynan, C.; Simoncelli, S.; Padilla-Parra, S.
Show abstract
We present Alpha-BET, a structure-guided strategy leveraging AlphaFold to identify optimal ALFA-tag insertion sites for minimally disruptive labeling of viral glycoproteins with high-affinity nanobodies. Applied to HIV-1 Env, SARS-CoV-2 S, and NiV G, Alpha-BET preserves structural integrity and function. For HIV-1 Env, we demonstrate super-resolution DNA-PAINT MINFLUX 3D imaging enabled by tag insertion, showcasing its power for visualizing native trimers in single virions and potential for broader applications in virus research.
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