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 1400-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 half-maximal inhibitory concentration (IC 50 ) of 4.0 nM (180 ng/mL). 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.
Bi-paratopic and multivalent VH domains block ACE2 binding and neutralize SARS-CoV-2
[저자] Colton J. Bracken, Shion A. Lim, Paige Solomon, Nicholas J. Rettko, Duy P. Nguyen, Beth Shoshana Zha, Kaitlin Schaefer, James R. Byrnes, Jie Zhou, Irene Lui, Jia Liu, Katarina Pance,
QCRG Structural Biology Consortium
Azumaya Caleigh M. Billesbølle Christian Brilot Axel F. Campbell Melody G. Diallo Amy Sasha Dickinson Miles Diwanji Devan Gupta Meghna Herrera Nadia Hoppe Nick Kratochvil Huong T. Liu Yanxin Merz Gregory E. Nguyen Henry C. Nowotny Carlos Moritz Michelle Owens Tristan W. Pourmal Sergei Puchades Cristina Rizo Alexandrea N. Schulze-Gahmen Ursula Smith Amber M. Sun Ming Titus Erron W. Young Iris D. Zhao Jianhua Asarnow Daniel Biel Justin Bowen Alisa Braxton Julian R. Bulkley David Chen Jen Chio Cynthia M. Seng Chio Un Deshpande Ishan Doan Loan Faust Bryan Flores Sebastian Jin Mingliang Kim Kate Lam Victor L. Li Fei Li Junrui Li Yen-Li Li Yang Liu Xi Lo Megan Lopez Kyle E. McKay Liam Melo Arthur A. Moss Frank R. III Nguyen Phuong Paulino Joana Ishwar Pawar Komal Peters Jessica K. Pospiech Thomas H. Jr. Safari Maliheh Sangwan Smriti Schaefer Kaitlin Tabios Mariano Thomas Paul V. Thwin Aye C. Trenker Raphael Trinidad Donovan Tse Eric Tsui Tsz Kin Martin Wang Feng Whitis Natalie Yu Zanlin Zhang Kaihua Zhang Yang Zhou Fengbo Rosenberg Oren S Verba Kliment A Agard David A Cheng Yifan Fraser James S Frost Adam Jura Natalia Kortemme Tanja Krogan Nevan J Manglik Aashish Southworth Daniel R. Stroud Robert M
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, Caleigh M. Azumaya, Christian Billesbølle, Axel F. Brilot, Melody G. Campbell, Amy Diallo, Miles Sasha Dickinson, Devan Diwanji, Meghna Gupta, Nadia Herrera, Nick Hoppe, Huong T. Kratochvil, Yanxin Liu, Gregory E. Merz, Henry C. Nguyen, Carlos Nowotny, Michelle Moritz, Tristan W. Owens, Sergei Pourmal, Cristina Puchades, Alexandrea N. Rizo, Ursula Schulze-Gahmen, Amber M. Smith, Ming Sun, Erron W. Titus, Iris D. Young, Jianhua Zhao, Daniel Asarnow, Justin Biel, Alisa Bowen, Julian R. Braxton, David Bulkley, Jen Chen, Cynthia M. Chio, Un Seng Chio, Ishan Deshpande, Loan Doan, Bryan Faust, Sebastian Flores, Mingliang Jin, Kate Kim, Victor L. Lam, Fei Li, Junrui Li, Yen-Li Li, Yang Li, Xi Liu, Megan Lo, Kyle E. Lopez, Liam McKay, Arthur A. Melo, Frank R. Moss, Phuong Nguyen, Joana Paulino, Komal Ishwar Pawar, Jessica K. Peters, Thomas H. Pospiech, Maliheh Safari, Smriti Sangwan, Kaitlin Schaefer, Mariano Tabios, Paul V. Thomas, Aye C. Thwin, Raphael Trenker, Donovan Trinidad, Eric Tse, Tsz Kin Martin Tsui, Feng Wang, Natalie Whitis, Zanlin Yu, Kaihua Zhang, Yang Zhang, Fengbo Zhou, Oren S Rosenberg, Kliment A Verba, David A Agard, Yifan Cheng, James S Fraser, Adam Frost, Natalia Jura, Tanja Kortemme, Nevan J Krogan, Aashish Manglik, Daniel R. Southworth, Robert M Stroud, Xin X. Zhou, Kevin K. Leung, James A. Wells
[Category] SARS,
[Article Type] Article
[Source] PMC
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