RT Journal Article
SR Electronic
T1 An ultra-high affinity synthetic nanobody blocks SARS-CoV-2 infection by locking Spike into an inactive conformation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.08.08.238469
DO 10.1101/2020.08.08.238469
A1 Michael Schoof
A1 Bryan Faust
A1 Reuben A. Saunders
A1 Smriti Sangwan
A1 Veronica Rezelj
A1 Nick Hoppe
A1 Morgane Boone
A1 Christian Bache Billesbølle
A1 Marcell Zimanyi
A1 Ishan Deshpande
A1 Jiahao Liang
A1 Aditya A. Anand
A1 Niv Dobzinski
A1 Beth Shoshana Zha
A1 Benjamin Barsi-Rhyne
A1 Vladislav Belyy
A1 Andrew W. Barile-Hill
A1 Sayan Gupta
A1 Camille R. Simoneau
A1 Kristoffer Leon
A1 Kris M. White
A1 Silke Nock
A1 Yuwei Liu
A1 Nevan J. Krogan
A1 Corie Y. Ralston
A1 Danielle L. Swaney
A1 Adolfo García-Sastre
A1 Melanie Ott
A1 Marco Vignuzzi
A1 QCRG Structural Biology Consortium
A1 Peter Walter
A1 Aashish Manglik
YR 2020
UL http://biorxiv.org/content/early/2020/08/10/2020.08.08.238469.abstract
AB Without an effective prophylactic solution, infections from SARS-CoV-2 continue to rise worldwide with devastating health and economic costs. SARS-CoV-2 gains entry into host cells via an interaction between its Spike protein and the host cell receptor angiotensin converting enzyme 2 (ACE2). Disruption of this interaction confers potent neutralization of viral entry, providing an avenue for vaccine design and for therapeutic antibodies. Here, we develop single-domain antibodies (nanobodies) that potently disrupt the interaction between the SARS-CoV-2 Spike and ACE2. By screening a yeast surface-displayed library of synthetic nanobody sequences, we identified a panel of nanobodies that bind to multiple epitopes on Spike and block ACE2 interaction via two distinct mechanisms. Cryogenic electron microscopy (cryo-EM) revealed that one exceptionally stable nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains (RBDs) locked into their inaccessible down-state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for SARS-CoV-2 Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains stability and function after aerosolization, lyophilization, and heat treatment. These properties may enable aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia, promising to yield a widely deployable, patient-friendly prophylactic and/or early infection therapeutic agent to stem the worst pandemic in a century.Competing Interest StatementM.Schoof, B.Faust, R.Saunders, N.Hoppe, P.Walter, and A.Manglik are inventors on a provisional patent describing anti-Spike nanobodies described in this manuscript.