Structure of the Inmazeb cocktail and resistance to Ebola virus escape

Creators: Rayaprolu, Vamseedhar; Fulton, Benjamin O.; Rafique, Ashique; Arturo, Emilia; Williams, Dewight; Hariharan, Chitra; Callaway, Heather; Parvate, Amar; Schendel, Sharon L.; Parekh, Diptiben; Hui, Sean; Shaffer, Kelly; Pascal, Kristen E.; Wloga, Elzbieta; Giordano, Stephanie; Negron, Nicole; Ni, Min; Copin, Richard; Atwal, Gurinder S.; Franklin, Matthew; Boytz, Ruth Mabel; Donahue, Callie; Davey, Robert; Baum, Alina; Kyratsous, Christos A.; Saphire, Erica Ollmann

Abstract

Monoclonal antibodies can provide important pre- or post-exposure protection against infectious disease for those not yet vaccinated or in individuals that fail to mount a protective immune response after vaccination. Inmazeb (REGN-EB3), a three-antibody cocktail against Ebola virus, lessened disease and improved survival in a controlled trial. Here, we present the cryo-EM structure at 3.1 Å of the Ebola virus glycoprotein, determined without symmetry averaging, in a simultaneous complex with the antibodies in the Inmazeb cocktail. This structure allows the modeling of previously disordered portions of the glycoprotein glycan cap, maps the non-overlapping epitopes of Inmazeb, and illuminates the basis for complementary activities and residues critical for resistance to escape by these and other clinically relevant antibodies. We further provide direct evidence that Inmazeb protects against the rapid emergence of escape mutants, whereas monotherapies even against conserved epitopes do not, supporting the benefit of a cocktail versus a monotherapy approach.

Additional Information

Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0). We gratefully acknowledge support from NIAID U19 AI142790, the Consortium for Immunotherapeutics against Emerging Viral Threats (E.O.S.), and the U.S. Department of Health and Health Services contract no. HHSO100201700016C (Regeneron). A portion of this project has been funded in part with federal funds from the Department of Health and Human Services, Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority, under OT number HHSO100201700020C. GM/CA@APS-23ID-B has been funded by the National Cancer Institute (ACB-12002) and the National Institute of General Medical Sciences (AGM-12006, P30GM138396). This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract no. DE-AC02-06CH11357. The Eiger 16M detector at GM/CA-XSD was funded by NIH grant S10 OD012289. Cryo-EM SPA datasets were collected at Arizona State University. We acknowledge the use of facilities within the Eyring Materials Center at Arizona State University supported in part by NNCI-ECCS-1542160. We would like to acknowledge Dr. Dewight Williams from the Eyring Materials Center at Arizona State University supported in part by NNCI-ECCS-1542160 for assisting in data collection. We thank all the staff of the NIH who supported this study, in particular, Kaleb Sharer, Russel Byrum, Jennifer Jackson, Sarah Klim, Danny Ragland, Marisa St Claire, and Lisa Hensley. The authors would like to thank Kristen Tramaglini for her continuous support with this project. Author contributions. Conceptualization, E.O.S., C.A.K., and A.B.; methodology, V.R., B.O.F., A.R., H.C., A.P., K.E.P., R.C., R.D., and E.W.; investigation, V.R., H.C., C.H., B.O.F., A.B., A.R., E.A., K.S., S.H., D.P., E.W., S.G., R.C., K.E.P., R.D., R.M.B., C.D., G.S.A., M.N., and N.N.; formal analysis, V.R., C.H., B.O.F., A.B., A.R., M.F., R.D., and K.E.P.; writing – original draft, V.R., E.O.S., B.O.F., and A.B.; writing – review & editing, V.R., E.O.S., B.O.F., M.F., S.L.S., A.P., C.A.K., and A.B.; visualization, V.R., B.O.F., A.R., and H.C.; supervision, E.O.S., C.A.K., M.F., and A.B.; resources, E.O.S., D.W., C.A.K., A.B., and A.R.; funding acquisition, E.O.S. and C.A.K. Data and code availability. The cryo-EM map has been deposited at the Electron Microscopy Data Bank (www.ebi.ac.uk/emdb). The accession number for the map is EMD-26005. The atomic model corresponding to the microscopy data has been deposited in the Protein Data Bank under the accession number PDB: 7TN9. No custom code was generated as part of the study. Any additional information required to reanalyze the data reported in this work paper is available from the Lead Contact upon request. Declaration of interests. The Regeneron employees have stock and/or options in the company. C.A.K. is an officer of the company. The antibodies are patented: WO/2016/123019. Inclusion and diversity. One or more of the authors of this paper self-identifies as a gender minority in their field of research. While citing references scientifically relevant for this work, we also actively worked to promote gender balance in our reference list.

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