@article{a3907e6646ff43dd8b5feef01cf531e9,
title = "Bi-paratopic and multivalent VH domains block ACE2 binding and neutralize SARS-CoV-2",
abstract = "Neutralizing agents against SARS-CoV-2 are urgently needed for the treatment and prophylaxis of COVID-19. Here, we present a strategy to rapidly identify and assemble synthetic human variable heavy (VH) domains toward neutralizing epitopes. We constructed a VH-phage library and targeted the angiotensin-converting enzyme 2 (ACE2) binding interface of the SARS-CoV-2 Spike receptor-binding domain (Spike-RBD). Using a masked selection approach, we identified VH binders to two non-overlapping epitopes and further assembled these into multivalent and bi-paratopic formats. These VH constructs showed increased affinity to Spike (up to 600-fold) and neutralization potency (up to 1,400-fold) on pseudotyped SARS-CoV-2 virus when compared to standalone VH domains. The most potent binder, a trivalent VH, neutralized authentic SARS-CoV-2 with a half-maximal inhibitory concentration (IC50) of 4.0 nM (180 ng ml−1). A cryo-EM structure of the trivalent VH bound to Spike shows each VH domain engaging an RBD at the ACE2 binding site, confirming our original design strategy. [Figure not available: see fulltext.]",
author = "{QCRG Structural Biology Consortium} and {CryoEM grid freezing/collection team} and {CryoEM data processing team} and {Mammalian cell expression team} and {Protein purification team} and {Crystallography team} and {Bacterial expression team} and {Infrastructure team} and {Leadership team} and Bracken, {Colton J.} and Lim, {Shion A.} and Paige Solomon and Rettko, {Nicholas J.} and Nguyen, {Duy P.} and Zha, {Beth Shoshana} and Kaitlin Schaefer and Byrnes, {James R.} and Jie Zhou and Irene Lui and Jia Liu and Katarina Pance and Azumaya, {Caleigh M.} and Braxton, {Julian R.} and Brilot, {Axel F.} and Meghna Gupta and Fei Li and Lopez, {Kyle E.} and Arthur Melo and Merz, {Gregory E.} and Frank Moss and Joana Paulino and Pospiech, {Thomas H.} and Sergei Pourmal and Cristina Puchades and Rizo, {Alexandrea N.} and Smith, {Amber M.} and Ming Sun and Thomas, {Paul V.} and Feng Wang and Zanlin Yu and Daniel Asarnow and Braxton, {Julian R.} and Campbell, {Melody G.} and Chio, {Cynthia M.} and Chio, {Un Seng} and Dickinson, {Miles Sasha} and Devan Diwanji and Bryan Faust and Nick Hoppe and Mingliang Jin and Fei Li and Junrui Li and Yanxin Liu and Merz, {Gregory E.} and Nguyen, {Henry C.} and Joana Paulino and Pospiech, {Thomas H.} and Sergei Pourmal and Krogan, {Nevan J.}",
note = "Publisher Copyright: {\textcopyright} 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2021",
month = jan,
doi = "10.1038/s41589-020-00679-1",
language = "English",
volume = "17",
pages = "113--121",
journal = "Nature Chemical Biology",
issn = "1552-4450",
publisher = "Nature Research",
number = "1",
}