TY - UNPB
T1 - Structure-based design of a highly stable, covalently-linked SARS-CoV-2 spike trimer with improved structural properties and immunogenicity
AU - Olmedillas, Eduardo
AU - Mann, Colin J.
AU - Peng, Weiwei
AU - Wang, Ying-Ting
AU - Avalos, Ruben Diaz
AU - Bedinger, Dan
AU - Valentine, Kristen
AU - Shafee, Norazizah
AU - Schendel, Sharon L.
AU - Yuan, Meng
AU - Lang, Guojun
AU - Rouet, Romain
AU - Christ, Daniel
AU - Jiang, Weidong
AU - Wilson, Ian A.
AU - Germann, Tim
AU - Shresta, Sujan
AU - Snijder, Joost
AU - Saphire, Erica Ollmann
PY - 2021/5/6
Y1 - 2021/5/6
N2 - The continued threat of SARS-CoV-2 to global health necessitates development of improved research tools and vaccines. We present an improved SARS-CoV-2 spike ectodomain, “VFLIP”, bearing five proline substitutions, a flexible cleavage site linker, and an inter-protomer disulfide bond. VFLIP displays significantly improved stability, high-yield production and retains its trimeric state without exogenous trimerization motifs. High-resolution cryo-EM and glycan profiling reveal that the VFLIP quaternary structure and glycosylation mimic the native spike on the viral surface. Further, VFLIP has enhanced affinity and binding kinetics relative to other stabilized spike proteins for antibodies in the Coronavirus Immunotherapeutic Consortium (CoVIC), and mice immunized with VFLIP exhibit potent neutralizing antibody responses against wild-type and B.1.351 live SARS-CoV-2. Taken together, VFLIP represents an improved tool for diagnostics, structural biology, antibody discovery, and vaccine design.Competing Interest StatementThe authors have declared no competing interest.
AB - The continued threat of SARS-CoV-2 to global health necessitates development of improved research tools and vaccines. We present an improved SARS-CoV-2 spike ectodomain, “VFLIP”, bearing five proline substitutions, a flexible cleavage site linker, and an inter-protomer disulfide bond. VFLIP displays significantly improved stability, high-yield production and retains its trimeric state without exogenous trimerization motifs. High-resolution cryo-EM and glycan profiling reveal that the VFLIP quaternary structure and glycosylation mimic the native spike on the viral surface. Further, VFLIP has enhanced affinity and binding kinetics relative to other stabilized spike proteins for antibodies in the Coronavirus Immunotherapeutic Consortium (CoVIC), and mice immunized with VFLIP exhibit potent neutralizing antibody responses against wild-type and B.1.351 live SARS-CoV-2. Taken together, VFLIP represents an improved tool for diagnostics, structural biology, antibody discovery, and vaccine design.Competing Interest StatementThe authors have declared no competing interest.
U2 - 10.1101/2021.05.06.441046
DO - 10.1101/2021.05.06.441046
M3 - Preprint
SP - 1
EP - 51
BT - Structure-based design of a highly stable, covalently-linked SARS-CoV-2 spike trimer with improved structural properties and immunogenicity
PB - bioRxiv
ER -
