@article{55703accdd55463d8c4662e89671c1be,
title = "Genetic and structural basis for SARS-CoV-2 variant neutralization by a two-antibody cocktail",
abstract = "Understanding the molecular basis for immune recognition of SARS-CoV-2 spike glycoprotein antigenic sites will inform the development of improved therapeutics. We determined the structures of two human monoclonal antibodies–AZD8895 and AZD1061–which form the basis of the investigational antibody cocktail AZD7442, in complex with the receptor-binding domain (RBD) of SARS-CoV-2 to define the genetic and structural basis of neutralization. AZD8895 forms an {\textquoteleft}aromatic cage{\textquoteright} at the heavy/light chain interface using germ line-encoded residues in complementarity-determining regions (CDRs) 2 and 3 of the heavy chain and CDRs 1 and 3 of the light chain. These structural features explain why highly similar antibodies (public clonotypes) have been isolated from multiple individuals. AZD1061 has an unusually long LCDR1; the HCDR3 makes interactions with the opposite face of the RBD from that of AZD8895. Using deep mutational scanning and neutralization escape selection experiments, we comprehensively mapped the crucial binding residues of both antibodies and identified positions of concern with regards to virus escape from antibody-mediated neutralization. Both AZD8895 and AZD1061 have strong neutralizing activity against SARS-CoV-2 and variants of concern with antigenic substitutions in the RBD. We conclude that germ line-encoded antibody features enable recognition of the SARS-CoV-2 spike RBD and demonstrate the utility of the cocktail AZD7442 in neutralizing emerging variant viruses.",
author = "Jinhui Dong and Zost, {Seth J.} and Greaney, {Allison J.} and Starr, {Tyler N.} and Dingens, {Adam S.} and Chen, {Elaine C.} and Chen, {Rita E.} and Case, {James Brett} and Sutton, {Rachel E.} and Pavlo Gilchuk and Jessica Rodriguez and Erica Armstrong and Christopher Gainza and Nargi, {Rachel S.} and Elad Binshtein and Xuping Xie and Xianwen Zhang and Shi, {Pei Yong} and James Logue and Stuart Weston and McGrath, {Marisa E.} and Frieman, {Matthew B.} and Tyler Brady and Tuffy, {Kevin M.} and Helen Bright and Loo, {Yueh Ming} and McTamney, {Patrick M.} and Esser, {Mark T.} and Carnahan, {Robert H.} and Diamond, {Michael S.} and Bloom, {Jesse D.} and Crowe, {James E.}",
note = "Funding Information: At the Fred Hutchinson Cancer Research Center, we thank A. Addetia for experimental assistance, the Flow Cytometry and Genomics core facilities and Scientific Computing supported by an ORIP grant no. S10OD028685. We thank A. Creanga and B. Graham of the U.S. National Institutes of Health (NIH) for the Vero-hACE2-TMPRSS2 cells. At AstraZeneca, we thank P. Warrener, C. Morehouse and D. Tabor for virus genome sequencing and spike variant analysis and K. Ren for the generation of protein reagents and related binding data. We thank B. Pinsky (Stanford) and M. Suthar (Emory) for providing the B.1.617.1 variant and R. Webby (St. Jude Children{\textquoteright}s Research Hospital) for the B.1.617.2 variant. Some schematics were created with Biorender.com. This work was supported by Defense Advanced Research Projects Agency grant nos. HR0011-18-2-0001 and HR0011-18-3-0001, NIH contract nos. 75N93019C00074 and 75N93019C00062, NIH grant nos. AI150739, AI157155, AI141707, AI083203, AI095202 and UL1TR001439, the Dolly Parton COVID-19 Research Fund at Vanderbilt, a grant from Fast Grants, Mercatus Center, George Mason University and funding from AstraZeneca. T.N.S. is a Washington Research Foundation Innovation Fellow at the University of Washington Institute for Protein Design and a Howard Hughes Medical Institute Fellow of the Damon Runyon Cancer Research Foundation (no. DRG-2381-19). J.E.C. is a recipient of the 2019 Future Insight Prize from Merck, which supported this work with a grant. J.D.B. is an investigator of the Howard Hughes Medical Institute. P.-Y.S. was supported by awards from the Sealy & Smith Foundation, Kleberg Foundation, John S. Dunn Foundation, Amon G. Carter Foundation, the Gilson Longenbaugh Foundation and the Summerfield Robert Foundation. J.B.C. is supported by a Helen Hay Whitney Foundation postdoctoral fellowship. X-ray diffraction data were collected at Beamline 21-ID-F and 21-ID-G at the Advanced Photon Source, a U.S. Department of Energy Office of Science User Facility operated for the Office of Science by Argonne National Laboratory under contract no. DE-AC02-06CH11357. Use of the LS-CAT Sector 21 was supported by the Michigan Economic Development Corporation and Michigan Technology Tri-Corridor (grant no. 085P1000817). Support for crystallography was provided from the Vanderbilt Center for Structural Biology. The content is solely the responsibility of the authors and does not necessarily represent the official views of the U.S. Government or the other sponsors. Funding Information: T.B., K.M.T., H.B., Y-M.L., P.M.M. and M.T.E. are employees of and may own stock in AstraZeneca. M.S.D. is a consultant for InBios International, Vir Biotechnology, Fortress Biotech and Carnival Corporation and on the scientific advisory boards of Moderna and Immunome. The Diamond laboratory has received funding support in sponsored research agreements from Moderna, Vir Biotechnology and Emergent BioSolutions. J.E.C. has served as a consultant for Luna Biologics, is a member of the scientific advisory board of Meissa Vaccines and is founder of IDBiologics. The Crowe laboratory at Vanderbilt University Medical Center has received sponsored research agreements from Takeda Vaccines, IDBiologics and AstraZeneca. Vanderbilt University has applied for patents concerning antibodies that are related to this work. Vanderbilt University has licensed certain rights to antibodies described in this paper to AstraZeneca. All other authors declare no competing interests. Publisher Copyright: {\textcopyright} 2021, The Author(s), under exclusive licence to Springer Nature Limited.",
year = "2021",
month = oct,
doi = "10.1038/s41564-021-00972-2",
language = "English",
volume = "6",
pages = "1233--1244",
journal = "Nature microbiology",
issn = "2058-5276",
number = "10",
}