Bispecific antibodies combine breadth, potency, and avidity of parental antibodies to neutralize sarbecoviruses

Laura Radić; Kwinten Sliepen; Victor Yin; Mitch Brinkkemper; Joan Capella-Pujol; Angela I Schriek; Jonathan L Torres; Sandhya Bangaru; Judith A Burger; Meliawati Poniman; Ilja Bontjer; Joey H Bouhuijs; David Gideonse; Dirk Eggink; Andrew B Ward; Albert J R Heck; Marit J Van Gils; Rogier W Sanders; Janke Schinkel

doi:10.1016/j.isci.2023.106540

Bispecific antibodies combine breadth, potency, and avidity of parental antibodies to neutralize sarbecoviruses

Laura Radić, Kwinten Sliepen, Victor Yin, Mitch Brinkkemper, Joan Capella-Pujol, Angela I Schriek, Jonathan L Torres, Sandhya Bangaru, Judith A Burger, Meliawati Poniman, Ilja Bontjer, Joey H Bouhuijs, David Gideonse, Dirk Eggink, Andrew B Ward, Albert J R Heck, Marit J Van Gils, Rogier W Sanders, Janke Schinkel

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

SARS-CoV-2 variants evade current monoclonal antibody therapies. Bispecific antibodies (bsAbs) combine the specificities of two distinct antibodies taking advantage of the avidity and synergy provided by targeting different epitopes. Here we used controlled Fab-arm exchange to produce bsAbs that neutralize SARS-CoV and SARS-CoV-2 variants, including Omicron and its subvariants, by combining potent SARS-CoV-2-specific neutralizing antibodies with broader antibodies that also neutralize SARS-CoV. We demonstrated that the parental antibodies rely on avidity for neutralization using bsAbs containing one irrelevant Fab arm. Using mass photometry to measure the formation of antibody:spike complexes, we determined that bsAbs increase binding stoichiometry compared to corresponding cocktails, without a loss of binding affinity. The heterogeneous binding pattern of bsAbs to spike, observed by negative-stain electron microscopy and mass photometry provided evidence for both intra- and inter-spike crosslinking. This study highlights the utility of cross-neutralizing antibodies for designing bivalent agents to combat circulating and future SARS-like coronaviruses.

Original language	English
Article number	106540
Pages (from-to)	1-23
Journal	iScience
Volume	26
Issue number	4
DOIs	https://doi.org/10.1016/j.isci.2023.106540
Publication status	Published - 21 Apr 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors

Funding

We thank Yoann Aldon and Tom Caniels for providing spike proteins and helpful discussions, Sylvie Koekkoek for help with cloning the antibodies, and Wouter Olijhoek and Jacqueline van Rijswijk for help with the pseudovirus neutralization assays. L.R. was funded by the AMC as part of the local scientific research incentive policy. J.S. is a recipient of a Vidi and Aspasia grant from the Netherlands Organization for Scientific Research ( NWO , grant numbers 91719372 and 015.015.042 ). R.W.S. is a recipient of a Vici grant from the Netherlands Organization for Scientific Research ( NWO , grant number 91818627 ). This research was supported by an Amsterdam institute for Infection and Immunity Postdoctoral grant (K.S.). A.J.R.H. acknowledges funding by the Netherlands Organization for Scientific Research ( NWO ) through the Spinoza Award SPI.2017.028. This work was supported by the Fondation Dormeur, Vaduz (R.W.S. and (M.J.v.G.).

Funders	Funder number
Amsterdam institute for Infection and Immunity Postdoctoral
Fondation Dormeur
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	015.015.042, 91818627, 91719372

Keywords

Immunology
Structural biology
Virology

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1-s2.0-S258900422300617X-mainFinal published version, 4.45 MBLicence: CC BY

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Radić, L., Sliepen, K., Yin, V., Brinkkemper, M., Capella-Pujol, J., Schriek, A. I., Torres, J. L., Bangaru, S., Burger, J. A., Poniman, M., Bontjer, I., Bouhuijs, J. H., Gideonse, D., Eggink, D., Ward, A. B., Heck, A. J. R., Van Gils, M. J., Sanders, R. W., & Schinkel, J. (2023). Bispecific antibodies combine breadth, potency, and avidity of parental antibodies to neutralize sarbecoviruses. iScience, 26(4), 1-23. Article 106540. https://doi.org/10.1016/j.isci.2023.106540

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doi = "10.1016/j.isci.2023.106540",

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Radić, L, Sliepen, K, Yin, V, Brinkkemper, M, Capella-Pujol, J, Schriek, AI, Torres, JL, Bangaru, S, Burger, JA, Poniman, M, Bontjer, I, Bouhuijs, JH, Gideonse, D, Eggink, D, Ward, AB, Heck, AJR, Van Gils, MJ, Sanders, RW & Schinkel, J 2023, 'Bispecific antibodies combine breadth, potency, and avidity of parental antibodies to neutralize sarbecoviruses', iScience, vol. 26, no. 4, 106540, pp. 1-23. https://doi.org/10.1016/j.isci.2023.106540

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AU - Poniman, Meliawati

AU - Bontjer, Ilja

AU - Bouhuijs, Joey H

AU - Gideonse, David

AU - Eggink, Dirk

AU - Ward, Andrew B

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AB - SARS-CoV-2 variants evade current monoclonal antibody therapies. Bispecific antibodies (bsAbs) combine the specificities of two distinct antibodies taking advantage of the avidity and synergy provided by targeting different epitopes. Here we used controlled Fab-arm exchange to produce bsAbs that neutralize SARS-CoV and SARS-CoV-2 variants, including Omicron and its subvariants, by combining potent SARS-CoV-2-specific neutralizing antibodies with broader antibodies that also neutralize SARS-CoV. We demonstrated that the parental antibodies rely on avidity for neutralization using bsAbs containing one irrelevant Fab arm. Using mass photometry to measure the formation of antibody:spike complexes, we determined that bsAbs increase binding stoichiometry compared to corresponding cocktails, without a loss of binding affinity. The heterogeneous binding pattern of bsAbs to spike, observed by negative-stain electron microscopy and mass photometry provided evidence for both intra- and inter-spike crosslinking. This study highlights the utility of cross-neutralizing antibodies for designing bivalent agents to combat circulating and future SARS-like coronaviruses.

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Bispecific antibodies combine breadth, potency, and avidity of parental antibodies to neutralize sarbecoviruses

Abstract

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Keywords

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