The smallest functional antibody fragment: Ultralong CDR H3 antibody knob regions potently neutralize SARS-CoV-2

Huang, Ruiqi; Warner Jenkins, Gabrielle; Kim, Yunjeong; Stanfield, Robyn L.; Singh, Amrinder; Martinez-Yamout, Maria; Kroon, Gerard J.; Torres, Jonathan L.; Jackson, Abigail M.; Kelley, Abigail; Shaabani, Namir; Zeng, Baisen; Bacica, Michael; Chen, Wen; Warner, Christopher; Radoicic, Jasmina; Joh, Joongho; Dinali Perera, Krishani; Sang, Huldah; Kim, Tae; Yao, Jianxiu; Zhao, Fangzhu; Sok, Devin; Burton, Dennis R.; Allen, Jeff; Harriman, William; Mwangi, Waithaka; Chung, Donghoon; Teijaro, John R.; Ward, Andrew B.; Dyson, H. Jane; Wright, Peter E.; Wilson, Ian A.; Chang, Kyeong-Ok; McGregor, Duncan; Smider, Vaughn V.

doi:10.1073/pnas.2303455120

The smallest functional antibody fragment: Ultralong CDR H3 antibody knob regions potently neutralize SARS-CoV-2

Journal Article · Mon Sep 18 00:00:00 EDT 2023 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.2303455120· OSTI ID:2423701

^[1]; ^[1]; ^[2]; ^[3]; Singh, Amrinder ^[3]; ^[3]; ^[3]; ^[3]; Jackson, Abigail M. ^[3]; Kelley, Abigail ^[1]; Shaabani, Namir ^[3]; Zeng, Baisen ^[4]; Bacica, Michael ^[5]; Chen, Wen ^[5]; Warner, Christopher ^[5]; Radoicic, Jasmina ^[5]; Joh, Joongho ^[6]; Dinali Perera, Krishani ^[2]; Sang, Huldah ^[2]; Kim, Tae ^[2] more »

Applied Biomedical Science Institute, San Diego, CA (United States)
Kansas State Univ., Manhattan, KS (United States)
The Scripps Research Inst., La Jolla, CA (United States)
OmniAb Inc., Emeryville, CA (United States)
Ligand Pharmaceuticals, San Diego, CA (United States)
Univ. of Louisville, KY (United States)
Applied Biomedical Science Institute, San Diego, CA (United States); The Scripps Research Inst., La Jolla, CA (United States)

Cows produce antibodies with a disulfide-bonded antigen-binding domain embedded within ultralong heavy chain third complementarity determining regions. This “knob” domain is analogous to natural cysteine-rich peptides such as knottins in that it is small and stable but can accommodate diverse loops and disulfide bonding patterns. We immunized cattle with SARS-CoV-2 spike and found ultralong CDR H3 antibodies that could neutralize several viral variants at picomolar IC₅₀ potencies in vitro and could protect from disease in vivo. The independent CDR H3 peptide knobs were expressed and maintained the properties of the parent antibodies. The knob interaction with SARS-CoV-2 spike was revealed by electron microscopy, X-ray crystallography, NMR spectroscopy, and mass spectrometry and established ultralong CDR H3-derived knobs as the smallest known recombinant independent antigen-binding fragment. Unlike other vertebrate antibody fragments, these knobs are not reliant on the immunoglobulin domain and have potential as a new class of therapeutics.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). Advanced Light Source (ALS); SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Sponsoring Organization:: Bill and Melinda Gates Foundation; National Institute of General Medical Sciences (NIGMS); National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231; AC02-06CH11357; AC02-76SF00515

OSTI ID:: 2423701

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 39 Vol. 120; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: English

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60 APPLIED LIFE SCIENCES
cow antibody
immunoglobulin
knob peptide
phage display
ultralong CDR3

The smallest functional antibody fragment: Ultralong CDR H3 antibody knob regions potently neutralize SARS-CoV-2

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