Bispecific IgG neutralizes SARS-CoV-2 variants and prevents escape in mice

Creators: De Gasparo, Raoul; Pedotti, Mattia; Simonelli, Luca; Nickl, Petr; Muecksch, Frauke; Cassaniti, Irene; Percivalle, Elena; Lorenzi, Julio C. C.; Mazzola, Federica; Magrì, Davide; Michalcikova, Tereza; Haviernik, Jan; Hönig, Václav; Mrazkova, Blanka; Polakova, Natalie; Fortova, Andrea; Tureckova, Jolana; Iatsiuk, Veronika; Di Girolamo, Salvatore; Palus, Martin; Zudova, Dagmar; Bednar, Petr; Bukova, Ivana; Bianchini, Filippo; Mehn, Dora; Nencka, Radim; Strakova, Petra; Pavlis, Oto; Rozman, Jan; Gioria, Sabrina; Camilla Sammartino, Josè; Giardina, Federica; Gaiarsa, Stefano; Pan-Hammarström, Qiang; Barnes, Christopher O.; Bjorkman, Pamela J.; Calzolai, Luigi; Piralla, Antonio; Baldanti, Fausto; Nussenzweig, Michel C.; Bieniasz, Paul D.; Hatziioannou, Theodora; Prochazka, Jan; Sedlacek, Radislav; Robbiani, Davide F.; Růžek, Daniel; Varani, Luca

Abstract

Neutralizing antibodies that target the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein are among the most promising approaches against COVID-19. A bispecific IgG1-like molecule (CoV-X2) has been developed on the basis of C121 and C135, two antibodies derived from donors who had recovered from COVID-19. Here we show that CoV-X2 simultaneously binds two independent sites on the RBD and, unlike its parental antibodies, prevents detectable spike binding to the cellular receptor of the virus, angiotensin-converting enzyme 2 (ACE2). Furthermore, CoV-X2 neutralizes wild-type SARS-CoV-2 and its variants of concern, as well as escape mutants generated by the parental monoclonal antibodies. We also found that in a mouse model of SARS-CoV-2 infection with lung inflammation, CoV-X2 protects mice from disease and suppresses viral escape. Thus, the simultaneous targeting of non-overlapping RBD epitopes by IgG-like bispecific antibodies is feasible and effective, and combines the advantages of antibody cocktails with those of single-molecule approaches.

Additional Information

© The Author(s), under exclusive licence to Springer Nature Limited 2021. Received 07 January 2021; Accepted 16 March 2021; Published 25 March 2021. D.F.R., L.V., Q.P.-H., F.Baldanti and L.C. have received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement no. 101003650. This work was also supported by SNF grant 31003A_182270 (L.V.); Lions Club Monteceneri (L.V.); George Mason University Fast Grant and IRB start-up funds (D.F.R.); NIH U01 AI151698 for the United World Antiviral Research Network (UWARN) (D.F.R. and M.C.N.); NIH grant P01-AI138398-S1 (M.C.N. and P.J.B.); 2U19AI111825 (M.C.N. and D.F.R.); the Caltech Merkin Institute for Translational Research and P50 AI150464 (P.J.B.); R37-AI64003 (P.D.B.); and R01AI78788 (T.H.); P.D.B. and M.C.N. are Howard Hughes Medical Institute Investigators. The study was also supported by the Czech Academy of Sciences and Czech Ministry of Agriculture (RVO 68378050 (R.S.) and RVO0518 (D.R.)); Czech Ministry of Education, Youth and Sports and the European Regional Development Fund (LM2018126; CZ.1.05/2.1.00/19.0395 and CZ.1.05/1.1.00/02.0109 (R.S.) and CZ.02.1.01/0.0/0.0/15_003/0000495 (D.R.)); Czech Science Foundation (20-14325S (D.R.)); the Bulgari Women & Science Fellowship in COVID-19 Research (F. Muecksch); the EU Joint Research Centre Exploratory Research program ('NanoMicrobials′; D. Magrì); and by Ricerca Finalizzata from Ministry of Health, Italy (grant no. GR-2013-02358399 (A.P.)). We are grateful for the high-performance computing resources that were provided by S. Bassini of CINECA to M. Hust, F. Bertoglio, F. Bognuda and E. Restivo. We thank V. Zatecka, V. Martinkova and L. Kutlikova for technical assistance; and V. Babak for help with statistical analyses. We are grateful to the late F. Diederich for their mentorship. Data availability: The data that support the findings of this study are available within the Article and its Supplementary Information. Any other data are available from the corresponding author upon reasonable request. Published data were taken from GenBank (https://www.ncbi.nlm.nih.gov/genbank/), UniProt (https://www.uniprot.org/), PDB (https://www.rcsb.org/) and the ViPR database (https://www.viprbrc.org/). Source data are provided with this paper. These authors contributed equally: Raoul De Gasparo, Mattia Pedotti. Author Contributions: R.D.G, M. Pedotti, L.S., F. Muecksch, J.C.C.L., F. Mazzola, D. Magrì, I.C., E.P., S.D.G., M. Palus, D. Mehn, S. Gioria, C.O.B., F. Bianchini, J.C.S., F.G. and S. Gaiarsa designed and carried out experiments and analysed results, and produced plasmids, antibodies and viral proteins. P.N., T.M., J.H., V.H, B.M., N.P., A.F., J.T., V.I., M. Palus, D.Z., P.B., I.B., P.S. and D.R., performed mouse experiments and analysed the results. L.V., D.F.R., D.R., Q.P.-H., F. Baldanti, A.P., L.C., P.J.B., M.C.N., P.D.B. and T.H. conceived and designed study and experiments, and analysed the results. P.N., T.M., R.N., O.P., J.P., J.R. and R.S. conceived and designed the mouse model. L.V., D.F.R., D.R. and R.D.G. wrote the manuscript, with input from all co-authors. Competing interests: The Institute for Research in Biomedicine has filed a provisional European patent application in connection with this work, on which L.V. is inventor (PCT/EP2020/085342). The Rockefeller University has filed a provisional US patent application (US 63/021,387) on coronavirus antibodies, on which D.F.R. and M.C.N. are inventors. Peer review information: Nature thanks Stanley Perlman and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

Errata

In this Article, authors Raoul De Gasparo and Mattia Pedotti should not be linked to affiliation 15, 'Howard Hughes Medical Institute, The Rockefeller University, New York, NY, USA' and should instead be linked to the equal contribution footnote, 16. In addition, affiliation 14 should read 'Università degli Studi di Pavia, Pavia, Italy'. The Article has been corrected online. De Gasparo, R., Pedotti, M., Simonelli, L. et al. Publisher Correction: Bispecific IgG neutralizes SARS-CoV-2 variants and prevents escape in mice. Nature (2021). https://doi.org/10.1038/s41586-021-03719-5

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Bispecific IgG neutralizes SARS-CoV-2 variants and prevents escape in mice

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