Structure-guided multivalent nanobodies block SARS-CoV-2 infection and suppress mutational escape
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Open Access
- 12 February 2021
- journal article
- research article
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 371 (6530), 691-+
- https://doi.org/10.1126/science.abe6230
Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic continues to spread with devastating consequences. For passive immunization efforts, nanobodies have size and cost advantages over conventional antibodies. Here, we generated four neutralizing nanobodies that target the receptor-binding domain of the SARS-CoV-2 spike protein. We defined two distinct binding epitopes using x-ray crystallography and cryo-electron microscopy. Based on the structures, we engineered multivalent nanobodies with more than 100-fold improved neutralizing activity than monovalent nanobodies. Biparatopic nanobody fusions suppressed the emergence of escape mutants. Several nanobody constructs neutralized through receptor-binding competition, while other monovalent and biparatopic nanobodies triggered aberrant activation of the spike fusion machinery. These premature conformational changes in the spike protein forestalled productive fusion, and rendered the virions non-infectious.Funding Information
- Bill and Melinda Gates Foundation (OPP1170236)
- University of Illinois at Urbana-Champaign
- Baden-Württemberg Stiftung (BWST_WSF-022)
- Horizon 2020 Framework Programme (CoroNAb)
- Universitätsklinikum Tübingen
- Deutsche Forschungsgemeinschaft (SFB1403-414786233)
- Deutsche Forschungsgemeinschaft (Germany’s Excellence Strategy – EXC2151 – 390873048)
- Deutsche Forschungsgemeinschaft (GE 976/9-2)
- Deutsche Forschungsgemeinschaft (SPP1923-429513120)
- Deutsche Forschungsgemeinschaft (SCHI1073)
- Deutsche Forschungsgemeinschaft (TRR237-369799452)
- Deutsche Forschungsgemeinschaft (SPP1923-429513120)
- Deutsche Forschungsgemeinschaft (322568668 (Emmy Noether Programme))
- Deutsche Forschungsgemeinschaft (Germany’s Excellence Strategy – EXC2151 – 390873048)
- Deutsche Forschungsgemeinschaft (GRK2168-272482170)
- Deutsche Forschungsgemeinschaft (TRR237-369799452)
- Deutsche Forschungsgemeinschaft (Germany’s Excellence Strategy – EXC2151 – 390873048)
- Deutsche Forschungsgemeinschaft (Germany’s Excellence Strategy – EXC2151 – 390873048)
- Knut och Alice Wallenbergs Stiftelse
- Vetenskapsrådet (2017-6702)
- Vetenskapsrådet (2018-3808)
- Ministerium für Wissenschaft, Forschung und Kunst Baden-Württemberg (33-7533-6-21/9/1)
- Klaus Tschira Stiftung (KT07)
- Bundesministerium für Bildung und Forschung (TTU 01.806)
This publication has 81 references indexed in Scilit:
- Site-specific C-terminal and internal loop labeling of proteins using sortase-mediated reactionsNature Protocols, 2013
- Different host cell proteases activate the SARS-coronavirus spike-protein for cell–cell and virus–cell fusionVirology, 2011
- Features and development of CootActa crystallographica. Section D, Structural biology, 2010
- PHENIX: a comprehensive Python-based system for macromolecular structure solutionActa crystallographica. Section D, Structural biology, 2010
- MolProbity: all-atom structure validation for macromolecular crystallographyActa crystallographica. Section D, Structural biology, 2009
- Jalview Version 2—a multiple sequence alignment editor and analysis workbenchBioinformatics, 2009
- Inference of Macromolecular Assemblies from Crystalline StateJournal of Molecular Biology, 2007
- Phasercrystallographic softwareJournal of Applied Crystallography, 2007
- Human Monoclonal Antibody Combination against SARS Coronavirus: Synergy and Coverage of Escape MutantsPLoS Medicine, 2006
- [20] Processing of X-ray diffraction data collected in oscillation modeMethods in Enzymology, 1997