Bispecific antibodies against the hepatitis C virus E1E2 envelope glycoprotein

Laura Radić; Anna Offersgaard; Tereza Kadavá; Ian Zon; Joan Capella-Pujol; Fabian Mulder; Sylvie Koekkoek; Vera Spek; Ana Chumbe; Jens Bukh; Marit J van Gils; Rogier W Sanders; Victor C Yin; Albert J R Heck; Judith M Gottwein; Kwinten Sliepen; Janke Schinkel

doi:10.1073/pnas.2420402122

Bispecific antibodies against the hepatitis C virus E1E2 envelope glycoprotein

Laura Radić, Anna Offersgaard, Tereza Kadavá, Ian Zon, Joan Capella-Pujol, Fabian Mulder, Sylvie Koekkoek, Vera Spek, Ana Chumbe, Jens Bukh, Marit J van Gils, Rogier W Sanders, Victor C Yin, Albert J R Heck, Judith M Gottwein, Kwinten Sliepen, Janke Schinkel^*

^*Corresponding author for this work

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

Abstract

Hepatitis C virus (HCV) currently causes about one million infections and 240,000 deaths worldwide each year. To reach the goal set by the World Health Organization of global HCV elimination by 2030, it is critical to develop a prophylactic vaccine. Broadly neutralizing antibodies (bNAbs) target the E1E2 envelope glycoproteins on the viral surface, can neutralize a broad range of the highly diverse circulating HCV strains, and are essential tools to inform vaccine design. However, bNAbs targeting a single E1E2 epitope might be limited in neutralization breadth, which can be enhanced by using combinations of bNAbs that target different envelope epitopes. We have generated 60 immunoglobulin G (IgG)-like bispecific antibodies (bsAbs) that can simultaneously target two distinct epitopes on E1E2. We combine non- or partially overlapping E1E2 specificities into three types of bsAbs, each containing a different hinge length. The majority of bsAbs shows retained or increased potency and breadth against a diverse panel of HCV pseudoparticles and HCV produced in cell culture compared to monospecific and cocktail controls. Additionally, we demonstrate that changes in the hinge length of bsAbs can alter the binding stoichiometry to E1E2. These results provide insights into the binding modes and the role of avidity in bivalent targeting of diverse E1E2 epitopes.This study illustrates how potential cooperative effects of HCV bNAbs can be utilized by strategically designing bispecific constructs. These HCV bsAbs can guide vaccine development and unlock novel therapeutic and prophylactic strategies against HCV and other (flavi)viruses.

Original language	English
Article number	e2420402122
Number of pages	12
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	122
Issue number	15
DOIs	https://doi.org/10.1073/pnas.2420402122
Publication status	Published - 7 Apr 2025

Bibliographical note

Publisher Copyright:
Copyright © 2025 the Author(s).

Funding

We thank Angela Schriek for providing the Fc\u03B3RIIIa-dimer protein and helping with the setup of the ELISA. We thank Andreas Chrysostomou for help with the statistical ranking analysis. We thank Steven de Taeye and Mitch Brinkkemper for helpful discussions. This work was supported by a Dutch Research Council (NWO, Dutch: Nederlandse Organisatie voor Wetenschappelijk Onderzoek) Vici Grant (no. 91818627) to R.W.S., by the Fondation Dormeur, Vaduz to M.J.v.G. and R.W.S., and by a Vidi and Aspasia grant from the NWO (grant numbers 91719372 and 015.015.042) to J.S. and by an Open Philanthropy grant to R.W.S., K.S., and J.S. Work on HCVcc was supported by grants from the Innovation Fund Denmark to J.M.G. and from the Novo Nordisk Foundation to J.B.

Funders	Funder number
Innovationsfonden
ELISA
Novo Nordisk Fonden
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	91818627
Fondation Dormeur	015.015.042, 91719372

Keywords

E1E2 envelope glycoprotein
bNAbs
bispecific antibodies
hepatitis C virus (HCV)
neutralization

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1073/pnas.2420402122Licence: CC BY-NC-ND

radić-et-al-bispecific-antibodies-against-the-hepatitis-c-virus-e1e2-envelope-glycoproteinFinal published version, 1.78 MBLicence: CC BY-NC-ND

Cite this

Radić, L., Offersgaard, A., Kadavá, T., Zon, I., Capella-Pujol, J., Mulder, F., Koekkoek, S., Spek, V., Chumbe, A., Bukh, J., van Gils, M. J., Sanders, R. W., Yin, V. C., Heck, A. J. R., Gottwein, J. M., Sliepen, K., & Schinkel, J. (2025). Bispecific antibodies against the hepatitis C virus E1E2 envelope glycoprotein. Proceedings of the National Academy of Sciences of the United States of America, 122(15), Article e2420402122. https://doi.org/10.1073/pnas.2420402122

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abstract = "Hepatitis C virus (HCV) currently causes about one million infections and 240,000 deaths worldwide each year. To reach the goal set by the World Health Organization of global HCV elimination by 2030, it is critical to develop a prophylactic vaccine. Broadly neutralizing antibodies (bNAbs) target the E1E2 envelope glycoproteins on the viral surface, can neutralize a broad range of the highly diverse circulating HCV strains, and are essential tools to inform vaccine design. However, bNAbs targeting a single E1E2 epitope might be limited in neutralization breadth, which can be enhanced by using combinations of bNAbs that target different envelope epitopes. We have generated 60 immunoglobulin G (IgG)-like bispecific antibodies (bsAbs) that can simultaneously target two distinct epitopes on E1E2. We combine non- or partially overlapping E1E2 specificities into three types of bsAbs, each containing a different hinge length. The majority of bsAbs shows retained or increased potency and breadth against a diverse panel of HCV pseudoparticles and HCV produced in cell culture compared to monospecific and cocktail controls. Additionally, we demonstrate that changes in the hinge length of bsAbs can alter the binding stoichiometry to E1E2. These results provide insights into the binding modes and the role of avidity in bivalent targeting of diverse E1E2 epitopes.This study illustrates how potential cooperative effects of HCV bNAbs can be utilized by strategically designing bispecific constructs. These HCV bsAbs can guide vaccine development and unlock novel therapeutic and prophylactic strategies against HCV and other (flavi)viruses.",

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Radić, L, Offersgaard, A, Kadavá, T, Zon, I, Capella-Pujol, J, Mulder, F, Koekkoek, S, Spek, V, Chumbe, A, Bukh, J, van Gils, MJ, Sanders, RW, Yin, VC, Heck, AJR, Gottwein, JM, Sliepen, K & Schinkel, J 2025, 'Bispecific antibodies against the hepatitis C virus E1E2 envelope glycoprotein', Proceedings of the National Academy of Sciences of the United States of America, vol. 122, no. 15, e2420402122. https://doi.org/10.1073/pnas.2420402122

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T1 - Bispecific antibodies against the hepatitis C virus E1E2 envelope glycoprotein

AU - Radić, Laura

AU - Offersgaard, Anna

AU - Kadavá, Tereza

AU - Zon, Ian

AU - Capella-Pujol, Joan

AU - Mulder, Fabian

AU - Koekkoek, Sylvie

AU - Spek, Vera

AU - Chumbe, Ana

AU - Bukh, Jens

AU - van Gils, Marit J

AU - Sanders, Rogier W

AU - Yin, Victor C

AU - Heck, Albert J R

AU - Gottwein, Judith M

AU - Sliepen, Kwinten

AU - Schinkel, Janke

PY - 2025/4/7

Y1 - 2025/4/7

N2 - Hepatitis C virus (HCV) currently causes about one million infections and 240,000 deaths worldwide each year. To reach the goal set by the World Health Organization of global HCV elimination by 2030, it is critical to develop a prophylactic vaccine. Broadly neutralizing antibodies (bNAbs) target the E1E2 envelope glycoproteins on the viral surface, can neutralize a broad range of the highly diverse circulating HCV strains, and are essential tools to inform vaccine design. However, bNAbs targeting a single E1E2 epitope might be limited in neutralization breadth, which can be enhanced by using combinations of bNAbs that target different envelope epitopes. We have generated 60 immunoglobulin G (IgG)-like bispecific antibodies (bsAbs) that can simultaneously target two distinct epitopes on E1E2. We combine non- or partially overlapping E1E2 specificities into three types of bsAbs, each containing a different hinge length. The majority of bsAbs shows retained or increased potency and breadth against a diverse panel of HCV pseudoparticles and HCV produced in cell culture compared to monospecific and cocktail controls. Additionally, we demonstrate that changes in the hinge length of bsAbs can alter the binding stoichiometry to E1E2. These results provide insights into the binding modes and the role of avidity in bivalent targeting of diverse E1E2 epitopes.This study illustrates how potential cooperative effects of HCV bNAbs can be utilized by strategically designing bispecific constructs. These HCV bsAbs can guide vaccine development and unlock novel therapeutic and prophylactic strategies against HCV and other (flavi)viruses.

AB - Hepatitis C virus (HCV) currently causes about one million infections and 240,000 deaths worldwide each year. To reach the goal set by the World Health Organization of global HCV elimination by 2030, it is critical to develop a prophylactic vaccine. Broadly neutralizing antibodies (bNAbs) target the E1E2 envelope glycoproteins on the viral surface, can neutralize a broad range of the highly diverse circulating HCV strains, and are essential tools to inform vaccine design. However, bNAbs targeting a single E1E2 epitope might be limited in neutralization breadth, which can be enhanced by using combinations of bNAbs that target different envelope epitopes. We have generated 60 immunoglobulin G (IgG)-like bispecific antibodies (bsAbs) that can simultaneously target two distinct epitopes on E1E2. We combine non- or partially overlapping E1E2 specificities into three types of bsAbs, each containing a different hinge length. The majority of bsAbs shows retained or increased potency and breadth against a diverse panel of HCV pseudoparticles and HCV produced in cell culture compared to monospecific and cocktail controls. Additionally, we demonstrate that changes in the hinge length of bsAbs can alter the binding stoichiometry to E1E2. These results provide insights into the binding modes and the role of avidity in bivalent targeting of diverse E1E2 epitopes.This study illustrates how potential cooperative effects of HCV bNAbs can be utilized by strategically designing bispecific constructs. These HCV bsAbs can guide vaccine development and unlock novel therapeutic and prophylactic strategies against HCV and other (flavi)viruses.

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KW - bispecific antibodies

KW - hepatitis C virus (HCV)

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Bispecific antibodies against the hepatitis C virus E1E2 envelope glycoprotein

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

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