Canonical Antibodies Adopt Distinct Binding Modes to Recognize Viral Glycan Shields
10 affiliations
- 1Vaccine Research Center National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda Maryland USA
- 2Cellular Biology Section Laboratory of Viral Diseases National Institute of Allergy and Infectious Diseases Bethesda Maryland USA
- 3Department of Biochemistry and Molecular Biophysics and Zuckerman Mind Brain Behavior Institute Columbia University New York New York USA
- 4Aaron Diamond AIDS Research Center Columbia University Vagelos College of Physicians and Surgeons New York New York USA
- 5Drukier Institute for Children's Health Department of Pediatrics Weill Cornell Medicine New York New York USA
- 6Department of Biology Johns Hopkins University Baltimore Maryland USA
- 7Vaccine Research Center Electron Microscopy Unit Cancer Research Technology Program Frederick National Laboratory For Cancer Research Sponsored By the National Cancer Institute Frederick Maryland USA
- 8Research Technologies Branch National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda Maryland USA
- 9Laboratory of Cell Biology National Cancer Institute National Institutes of Health Bethesda Maryland USA
- 10Laboratory of Immunoregulation National Institute of Allergy and Infectious Diseases National Institutes of Health Bethesda Maryland USA
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.
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BibTeX
@article{cheng2026canonical,
title = {{Canonical Antibodies Adopt Distinct Binding Modes to Recognize Viral Glycan Shields}},
author = {Jiaxuan Cheng and Evan M. Cale and Amirabbas Maghsoudi and Matthew S. Sutton and Nancy S. Longo and Rebecca A. Gillespie and Lingshu Wang and Ivan Kosik and Sue Chong and Atsuhiro Yasuhara and Tatsiana Bylund and Arne Schön and Prabhanshu Tripathi and Yaroslav Tsybovsky and Haotian Lei and Nicholas C. Morano and Allison B. Lupatkin and Abraham J. Morton and Zabrina C. Lang and Jordan E. Becker and Isabella R. Frascilla and Myungjin Lee and Ning Li and Cuiping Liu and Ryan S. Roark and Chen‐Hsiang Shen and I‐Ting Teng and David J. Van Wazer and Danyi Wang and Lingyuan Wu and Goran Ahlsen and Mike Castro and Haijuan Du and Michael J. Ernandes and Tiansheng Li and Bob C. Lin and Mark K. Louder and Krisha McKee and Jonah S. Merriam and Sijy O'Dell and Li Ou and Sergei Pletnev and David Prikryl and Qi Qiu and Sarah Rubin and Mallika Sastry and Stephen D. Schmidt and Asif Shajahan and Andrea R. Shiakolas and Sanjay Srivatsan and Baoshan Zhang and Qiong Zhou and Mark Connors and Jason G. Gall and Yicheng Guo and Rick K. Huang and Yaoxing Huang and Richard A. Koup and Q. Paula Lei and John R. Mascola},
journal = {Advanced Science},
year = {2026},
month = aug,
doi = {10.1002/advs.77172},
url = {https://doi.org/10.1002/advs.77172},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Canonical Antibodies Adopt Distinct Binding Modes to Recognize Viral Glycan Shields
AU - Jiaxuan Cheng
AU - Evan M. Cale
AU - Amirabbas Maghsoudi
AU - Matthew S. Sutton
AU - Nancy S. Longo
AU - Rebecca A. Gillespie
AU - Lingshu Wang
AU - Ivan Kosik
AU - Sue Chong
AU - Atsuhiro Yasuhara
AU - Tatsiana Bylund
AU - Arne Schön
AU - Prabhanshu Tripathi
AU - Yaroslav Tsybovsky
AU - Haotian Lei
AU - Nicholas C. Morano
AU - Allison B. Lupatkin
AU - Abraham J. Morton
AU - Zabrina C. Lang
AU - Jordan E. Becker
AU - Isabella R. Frascilla
AU - Myungjin Lee
AU - Ning Li
AU - Cuiping Liu
AU - Ryan S. Roark
AU - Chen‐Hsiang Shen
AU - I‐Ting Teng
AU - David J. Van Wazer
AU - Danyi Wang
AU - Lingyuan Wu
AU - Goran Ahlsen
AU - Mike Castro
AU - Haijuan Du
AU - Michael J. Ernandes
AU - Tiansheng Li
AU - Bob C. Lin
AU - Mark K. Louder
AU - Krisha McKee
AU - Jonah S. Merriam
AU - Sijy O'Dell
AU - Li Ou
AU - Sergei Pletnev
AU - David Prikryl
AU - Qi Qiu
AU - Sarah Rubin
AU - Mallika Sastry
AU - Stephen D. Schmidt
AU - Asif Shajahan
AU - Andrea R. Shiakolas
AU - Sanjay Srivatsan
AU - Baoshan Zhang
AU - Qiong Zhou
AU - Mark Connors
AU - Jason G. Gall
AU - Yicheng Guo
AU - Rick K. Huang
AU - Yaoxing Huang
AU - Richard A. Koup
AU - Q. Paula Lei
AU - John R. Mascola
PY - 2026
DA - 2026/08/14
JO - Advanced Science
DO - 10.1002/advs.77172
UR - https://doi.org/10.1002/advs.77172
AB - 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.
SN - 2198-3844
ER -