Canonical Antibodies Adopt Distinct Binding Modes to Recognize Viral Glycan Shields

Viral entry glycoproteins are often shielded from immune recognition by dense N-linked glycans that limit antibody access to protein epitopes. While glycan-reactive antibodies with unusual architectures have been described, how canonical Y-shaped antibodies engage these glycan-rich surfaces remains poorly defined. Here, we characterize two human antibodies, VRC35 and VRC36, isolated from an HIV-1-infected donor, that recognize diverse glycosylated viral glycoproteins. Cryo-electron microscopy structural analyses of these antibodies in complex with viral entry glycoproteins, including HIV-1 envelope, influenza hemagglutinin, SARS-CoV-2 spike, and the Lassa virus glycoprotein complex, reveal adaptive Fab stoichiometries ranging from single-Fab binding to dimeric and higher-order assemblies are mediated by intra- and inter-IgG interactions that depend on local glycan organization. Dense glycan clustering on HIV-1 and influenza glycoproteins supports multivalent Fab assemblies and correlates with neutralization activity, whereas sparse glycan environments on SARS-CoV-2 and Lassa virus favor weak or heterogeneous engagement without neutralization. Structural and mutational analyses further demonstrate that homotypic Fab-Fab interactions stabilize multivalent engagement and contribute to neutralizing activity. Together, these findings define a structural framework in which viral glycan organization constrains antibody valency and engagement, while somatic hypermutation contributes to the acquisition of homotypic Fab-Fab interactions that facilitate multivalent recognition of viral glycan shields.

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Publication Details

Journal
Advanced Science
Published
2026-08-14
DOI
https://doi.org/10.1002/advs.77172
Primary Topic
HIV Research and Treatment
Type
article
Field-Weighted Citation Impact
0.00

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article

Canonical Antibodies Adopt Distinct Binding Modes to Recognize Viral Glycan Shields

Michael J. Ernandes, Ivan Košík, Tongqing Zhou, Andrea R. Shiakolas et al.
Advanced Science
HIV Research and Treatment
article

Canonical Antibodies Adopt Distinct Binding Modes to Recognize Viral Glycan Shields

Michael J. Ernandes, Ivan Košík, Tongqing Zhou, Andrea R. Shiakolas, David Přikryl, Jiaxuan Cheng, Danyi Wang, Patrick C. Wilson, Arne Schön, Bob C. Lin, Tiansheng Li, Atsuhiro Yasuhara, Stephen D. Schmidt, Nancy S. Longo, David J. Van Wazer, Tatsiana Bylund, Myungjin Lee, Mark Connors, Qi Qiu, Lingyuan Wu, Yicheng Guo, Peter D. Kwong, Göran Ahlsén, Cuiping Liu, Masaru Kanekiyo, Yaoxing Huang, Matthew S. Sutton, Mike Castro, Mark K. Louder, Amirabbas Maghsoudi, Lawrence Shapiro, Q. Paula Lei, Prabhanshu Tripathi, Ryan S. Roark, Chen‐Hsiang Shen, Rebecca A. Gillespie, Jason G. Gall, Zizhang Sheng, Asif Shajahan, Sanjay Srivatsan, Nicole A. Doria‐Rose, I‐Ting Teng, Reda Rawi, Jonah S. Merriam, Qiong Zhou, Li Ou, Jordan E. Becker, A. Morton, J W Yewdell, Mallika Sastry, Krisha McKee, Haijuan Du, Leonid Serebryannyy, Sue Chong, Allison B. Lupatkin, Rick K. Huang, Sergei Pletnev, Lingshu Wang, Sarah Rubin, Haotian Lei, Baoshan Zhang, Sijy O'Dell, Yaroslav Tsybovsky, John R. Mascola, Zabrina C. Lang, Richard A. Koup, Nicole A. Doria‐Rose, Ning Li, Evan M. Cale, Isabella R. Frascilla, Theodore C. Pierson, David D. Ho
article en

Abstract

Viral entry glycoproteins are often shielded from immune recognition by dense N-linked glycans that limit antibody access to protein epitopes. While glycan-reactive antibodies with unusual architectures have been described, how canonical Y-shaped antibodies engage these glycan-rich surfaces remains poorly defined. Here, we characterize two human antibodies, VRC35 and VRC36, isolated from an HIV-1-infected donor, that recognize diverse glycosylated viral glycoproteins. Cryo-electron microscopy structural analyses of these antibodies in complex with viral entry glycoproteins, including HIV-1 envelope, influenza hemagglutinin, SARS-CoV-2 spike, and the Lassa virus glycoprotein complex, reveal adaptive Fab stoichiometries ranging from single-Fab binding to dimeric and higher-order assemblies are mediated by intra- and inter-IgG interactions that depend on local glycan organization. Dense glycan clustering on HIV-1 and influenza glycoproteins supports multivalent Fab assemblies and correlates with neutralization activity, whereas sparse glycan environments on SARS-CoV-2 and Lassa virus favor weak or heterogeneous engagement without neutralization. Structural and mutational analyses further demonstrate that homotypic Fab-Fab interactions stabilize multivalent engagement and contribute to neutralizing activity. Together, these findings define a structural framework in which viral glycan organization constrains antibody valency and engagement, while somatic hypermutation contributes to the acquisition of homotypic Fab-Fab interactions that facilitate multivalent recognition of viral glycan shields.

Advanced Science
National Institutes of Health (US), Allen Institute for Brain Science (US), Johns Hopkins University (US), Aaron Diamond AIDS Research Center (US), Cornell University (US), Children's Cancer and Blood Foundation (US), Frederick National Laboratory for Cancer Research (US), National Institute of Allergy and Infectious Diseases (US), National Cancer Institute (US)
Frederick National Laboratory for Cancer Research
Good health and well-being
Openalex Percentile: Top 11%
HIV Research and Treatment
DOIOpen Access PDFOpenAlex
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