@article{1b4e85a9cffb499095138dae9e92b10a,
title = "Structure-guided multivalent nanobodies block SARS-CoV-2 infection and suppress mutational escape",
abstract = "The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread, with devastating consequences. For passive immunization efforts, nanobodies have size and cost advantages over conventional antibodies. In this study, we generated four neutralizing nanobodies that target the receptor binding domain of the SARS-CoV-2 spike protein. We used x-ray crystallography and cryo-electron microscopy to define two distinct binding epitopes. On the basis of these structures, we engineered multivalent nanobodies with more than 100 times the neutralizing activity of monovalent nanobodies. Biparatopic nanobody fusions suppressed the emergence of escape mutants. Several nanobody constructs neutralized through receptor binding competition, whereas 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 noninfectious.",
keywords = "Novel coronavirus, COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Pandemic, 2019-nCoV",
author = "Paul-albert Koenig and Hrishikesh Das and Hejun Liu and K{\"u}mmerer, {Beate M.} and Gohr, {Florian N.} and Lea-marie Jenster and Schiffelers, {Lisa D. J.} and Tesfamariam, {Yonas M.} and Miki Uchima and Wuerth, {Jennifer D.} and Karl Gatterdam and Natalia Ruetalo and Christensen, {Maria H.} and Fandrey, {Caroline I.} and Sabine Normann and T{\"o}dtmann, {Jan M. P.} and Steffen Pritzl and Leo Hanke and Jannik Boos and Meng Yuan and Xueyong Zhu and Schmid-burgk, {Jonathan L.} and Hiroki Kato and Michael Schindler and Wilson, {Ian A.} and Matthias Geyer and Ludwig, {Kerstin U.} and H{\"a}llberg, {B. Martin} and Wu, {Nicholas C.} and Schmidt, {Florian I.}",
note = "Funding Information: We thank J. Ciftcioglu, G. Saavedra Perez Salas, J. Wieseler, and G. V. Gese for technical support; R. Businger for help with data analysis; G. Horvath and the Microscopy Core Facility of the Medical Faculty at the University of Bonn for providing help and services; the Karolinska Institutet for access to their 3D-EM facility, which was used for collection of all cryo-EM data; H. Tien for technical support with the crystallization robot; R. Stanfield for assistance with x-ray data collection; and the staff of Stanford Synchrotron Radiation Lightsource (SSRL) beamline 12-1 and Advanced Photon Source (APS) beamline 23ID-B for assistance. This study was partly supported by the Deutsche Forschungsgemeinschaft (DFG; German Research Foundation): GRK2168-272482170 (F.I.S.), SFB1403-414786233 (F.I.S.), TRR237-369799452 (F.I.S. and B.M.K.), SPP1923-429513120 (F.I.S. and M.S.), the Emmy Noether Programme 322568668 (F.I.S.) and LU 1933-3/1 (K.U.L.), GE 976/9-2 (M.G.), SCHI1073 (M.S.), and Germany's Excellence Strategy-EXC2151-390873048 (F.I.S., J.L.S.-B., H.K., and M.G.); by funds from the Medical Faculty of the University of Bonn (P.-A.K. and F.I.S.); by the Klaus Tschira Boost Fund (KT07) (F.I.S.), the Bundesministerium f?r Bildung und Forschung (BMBF), project TTU 01.806 (B.M.K.); by the Baden-W?rttemberg foundation (BW-Stiftung, BWST_WSF-022) (M.S.), the MWK Baden-W?rttemberg (33-7533-6-21/9/1) (M.S.), the University Hospital T?bingen (M.S.); the European Union's Horizon 2020 research and innovation program (CoroNAb consortium, L.H.); the Swedish Research Council (2017-6702 and 2018-3808) (B.M.H.); the Knut and Alice Wallenberg Foundation (B.M.H.); startup funds from the University of Illinois at Urbana-Champaign (N.C.W.); and the Bill and Melinda Gates Foundation OPP1170236 and INV-004923 (I.A.W.). Use of the SSRL, SLAC National Accelerator Laboratory, is supported by the U.S. Department of Energy (DOE), Office of Science, Office of Basic Energy Sciences, under contract DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research and by the National Institutes of Health, National Institute of General Medical Sciences (NIGMS) (including P41GM103393). General Medical Sciences and Cancer Institutes Structural Biology Facility at the Advanced Photon Source (APS) has been funded by federal funds from the National Cancer Institute (ACB-12002) and the NIGMS (AGM-12006). This research used resources of the APS, a DOE Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract DE-AC02-06CH11357. Extraordinary facility operations were supported in part by the DOE Office of Science through the National Virtual Biotechnology Laboratory, a consortium of DOE national laboratories focused on the response to COVID-19 with funding provided by the Coronavirus CARES Act. Publisher Copyright: Copyright {\textcopyright} 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",
year = "2021",
month = feb,
day = "12",
doi = "10.1126/science.abe6230",
language = "English (US)",
volume = "371",
pages = "eabe6230",
journal = "Science",
issn = "0036-8075",
publisher = "American Association for the Advancement of Science",
number = "6530",
}