Probing Affinity, Avidity, Anticooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses

Victor Yin; Szu-Hsueh Lai; Tom G Caniels; Philip J M Brouwer; Mitch Brinkkemper; Yoann Aldon; Hejun Liu; Meng Yuan; Ian A Wilson; Rogier W Sanders; Marit J van Gils; Albert J R Heck

doi:10.1021/acscentsci.1c00804

Probing Affinity, Avidity, Anticooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses

Victor Yin, Szu-Hsueh Lai, Tom G Caniels, Philip J M Brouwer, Mitch Brinkkemper, Yoann Aldon, Hejun Liu, Meng Yuan, Ian A Wilson, Rogier W Sanders, Marit J van Gils, Albert J R Heck

University of Amsterdam, VU University Amsterdam
Department of Integrative Structural and Computational Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA; Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. Electronic address: [email protected].

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

Abstract

Determining how antibodies interact with the spike (S) protein of the SARS-CoV-2 virus is critical for combating COVID-19. Structural studies typically employ simplified, truncated constructs that may not fully recapitulate the behavior of the original complexes. Here, we combine two single particle mass analysis techniques (mass photometry and charge-detection mass spectrometry) to enable the measurement of full IgG binding to the trimeric SARS-CoV-2 S ectodomain. Our experiments reveal that antibodies targeting the S-trimer typically prefer stoichiometries lower than the symmetry-predicted 3:1 binding. We determine that this behavior arises from the interplay of steric clashes and avidity effects that are not reflected in common antibody constructs (i.e., Fabs). Surprisingly, these substoichiometric complexes are fully effective at blocking ACE2 binding despite containing free receptor binding sites. Our results highlight the importance of studying antibody/antigen interactions using complete, multimeric constructs and showcase the utility of single particle mass analyses in unraveling these complex interactions.

Original language	English
Pages (from-to)	1863-1873
Number of pages	11
Journal	ACS Central Science
Volume	7
Issue number	11
DOIs	https://doi.org/10.1021/acscentsci.1c00804
Publication status	Published - 24 Nov 2021

Bibliographical note

Funding Information:
This research received funding through the Dutch Research Council (NWO) funding The Netherlands Proteomics Centre through the X-omics Road Map program (project 184.034.019). A.J.R.H. acknowledges support from The Netherlands Organization for Scientific Research (NWO) through the Spinoza Award SPI.2017.028 to A.J.R.H. R.W.S. acknowledges support from The Netherlands Organization for Scientific Research (NWO) through a Vici grant. The authors also acknowledge support from the Bill & Melinda Gates Foundation grants INV-002022 and INV-008818 (to R.W.S.), INV-024617 (M.J.v.G.), and INV-004923 (to I.A.W.). M.J.v.G. is a recipient of an Amsterdam UMC AMC Fellowship.

Publisher Copyright:
© 2021 The Authors. Published by American Chemical Society.

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Yin, V., Lai, S.-H., Caniels, T. G., Brouwer, P. J. M., Brinkkemper, M., Aldon, Y., Liu, H., Yuan, M., Wilson, I. A., Sanders, R. W., van Gils, M. J., & Heck, A. J. R. (2021). Probing Affinity, Avidity, Anticooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses. ACS Central Science, 7(11), 1863-1873. https://doi.org/10.1021/acscentsci.1c00804

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title = "Probing Affinity, Avidity, Anticooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses",

abstract = "Determining how antibodies interact with the spike (S) protein of the SARS-CoV-2 virus is critical for combating COVID-19. Structural studies typically employ simplified, truncated constructs that may not fully recapitulate the behavior of the original complexes. Here, we combine two single particle mass analysis techniques (mass photometry and charge-detection mass spectrometry) to enable the measurement of full IgG binding to the trimeric SARS-CoV-2 S ectodomain. Our experiments reveal that antibodies targeting the S-trimer typically prefer stoichiometries lower than the symmetry-predicted 3:1 binding. We determine that this behavior arises from the interplay of steric clashes and avidity effects that are not reflected in common antibody constructs (i.e., Fabs). Surprisingly, these substoichiometric complexes are fully effective at blocking ACE2 binding despite containing free receptor binding sites. Our results highlight the importance of studying antibody/antigen interactions using complete, multimeric constructs and showcase the utility of single particle mass analyses in unraveling these complex interactions.",

author = "Victor Yin and Szu-Hsueh Lai and Caniels, {Tom G} and Brouwer, {Philip J M} and Mitch Brinkkemper and Yoann Aldon and Hejun Liu and Meng Yuan and Wilson, {Ian A} and Sanders, {Rogier W} and {van Gils}, {Marit J} and Heck, {Albert J R}",

note = "Funding Information: This research received funding through the Dutch Research Council (NWO) funding The Netherlands Proteomics Centre through the X-omics Road Map program (project 184.034.019). A.J.R.H. acknowledges support from The Netherlands Organization for Scientific Research (NWO) through the Spinoza Award SPI.2017.028 to A.J.R.H. R.W.S. acknowledges support from The Netherlands Organization for Scientific Research (NWO) through a Vici grant. The authors also acknowledge support from the Bill & Melinda Gates Foundation grants INV-002022 and INV-008818 (to R.W.S.), INV-024617 (M.J.v.G.), and INV-004923 (to I.A.W.). M.J.v.G. is a recipient of an Amsterdam UMC AMC Fellowship. Publisher Copyright: {\textcopyright} 2021 The Authors. Published by American Chemical Society.",

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Yin, V, Lai, S-H, Caniels, TG, Brouwer, PJM, Brinkkemper, M, Aldon, Y, Liu, H, Yuan, M, Wilson, IA, Sanders, RW, van Gils, MJ & Heck, AJR 2021, 'Probing Affinity, Avidity, Anticooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses', ACS Central Science, vol. 7, no. 11, pp. 1863-1873. https://doi.org/10.1021/acscentsci.1c00804

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AU - Yin, Victor

AU - Lai, Szu-Hsueh

AU - Caniels, Tom G

AU - Brouwer, Philip J M

AU - Brinkkemper, Mitch

AU - Aldon, Yoann

AU - Liu, Hejun

AU - Yuan, Meng

AU - Wilson, Ian A

AU - Sanders, Rogier W

AU - van Gils, Marit J

AU - Heck, Albert J R

N1 - Funding Information: This research received funding through the Dutch Research Council (NWO) funding The Netherlands Proteomics Centre through the X-omics Road Map program (project 184.034.019). A.J.R.H. acknowledges support from The Netherlands Organization for Scientific Research (NWO) through the Spinoza Award SPI.2017.028 to A.J.R.H. R.W.S. acknowledges support from The Netherlands Organization for Scientific Research (NWO) through a Vici grant. The authors also acknowledge support from the Bill & Melinda Gates Foundation grants INV-002022 and INV-008818 (to R.W.S.), INV-024617 (M.J.v.G.), and INV-004923 (to I.A.W.). M.J.v.G. is a recipient of an Amsterdam UMC AMC Fellowship. Publisher Copyright: © 2021 The Authors. Published by American Chemical Society.

PY - 2021/11/24

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N2 - Determining how antibodies interact with the spike (S) protein of the SARS-CoV-2 virus is critical for combating COVID-19. Structural studies typically employ simplified, truncated constructs that may not fully recapitulate the behavior of the original complexes. Here, we combine two single particle mass analysis techniques (mass photometry and charge-detection mass spectrometry) to enable the measurement of full IgG binding to the trimeric SARS-CoV-2 S ectodomain. Our experiments reveal that antibodies targeting the S-trimer typically prefer stoichiometries lower than the symmetry-predicted 3:1 binding. We determine that this behavior arises from the interplay of steric clashes and avidity effects that are not reflected in common antibody constructs (i.e., Fabs). Surprisingly, these substoichiometric complexes are fully effective at blocking ACE2 binding despite containing free receptor binding sites. Our results highlight the importance of studying antibody/antigen interactions using complete, multimeric constructs and showcase the utility of single particle mass analyses in unraveling these complex interactions.

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Probing Affinity, Avidity, Anticooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses

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