학술논문
Bi-paratopic and multivalent VH domains block ACE2 binding and neutralize SARS-CoV-2
Document Type
article
Author
Azumaya, Caleigh M; Braxton, Julian R; Brilot, Axel F; Gupta, Meghna; Li, Fei; Lopez, Kyle E; Melo, Arthur; Merz, Gregory E; Moss, Frank; Paulino, Joana; Pospiech, Thomas H; Pourmal, Sergei; Puchades, Cristina; Rizo, Alexandrea N; Smith, Amber M; Sun, Ming; Thomas, Paul V; Wang, Feng; Yu, Zanlin; Asarnow, Daniel; Campbell, Melody G; Chio, Cynthia M; Chio, Un Seng; Dickinson, Miles Sasha; Diwanji, Devan; Faust, Bryan; Hoppe, Nick; Jin, Mingliang; Li, Junrui; Liu, Yanxin; Nguyen, Henry C; Sangwan, Smriti; Trenker, Raphael; Trinidad, Donovan; Tse, Eric; Zhang, Kaihua; Zhou, Fengbo; Billesboelle, Christian; Bowen, Alisa; Li, Yen-Li; Nguyen, Phuong; Nowotny, Carlos; Safari, Mali; Schaefer, Kaitlin; Tsui, Tsz Kin Martin; Whitis, Natalie; Zhao, Jianhua; Kim, Kate; Moritz, Michelle; Owens, Tristan W; Peters, Jessica K; Biel, Justin; Deshpande, Ishan; Herrera, Nadia; Kratochvil, Huong T; Liu, Xi; Schulze-Gahmen, Ursula; Young, Iris D; Chen, Jen; Diallo, Amy; Doan, Loan; Flores, Sebastian; Lam, Victor L; Li, Yang; Lo, Megan; Thwin, Aye C; Titus, Erron W; Zhang, Yang; Bulkley, David
Source
Nature Chemical Biology. 17(1)
Subject
Language
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.