PT  - JOURNAL ARTICLE
AU  - Eduardo Olmedillas
AU  - Colin J. Mann
AU  - Weiwei Peng
AU  - Ying-Ting Wang
AU  - Ruben Diaz Avalos
AU  - Dan Bedinger
AU  - Kristen Valentine
AU  - Norazizah Shafee
AU  - Sharon L. Schendel
AU  - Meng Yuan
AU  - Guojun Lang
AU  - Romain Rouet
AU  - Daniel Christ
AU  - Weidong Jiang
AU  - Ian A. Wilson
AU  - Tim Germann
AU  - Sujan Shresta
AU  - Joost Snijder
AU  - Erica Ollmann Saphire
TI  - Structure-based design of a highly stable, covalently-linked SARS-CoV-2 spike trimer with improved structural properties and immunogenicity
AID  - 10.1101/2021.05.06.441046
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.05.06.441046
4099  - http://biorxiv.org/content/early/2021/05/06/2021.05.06.441046.short
4100  - http://biorxiv.org/content/early/2021/05/06/2021.05.06.441046.full
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.