A cell-free nanobody engineering platform rapidly generates SARS-CoV-2 neutralizing nanobodies
Name
s41467-021-25777-z.pdf
Description
Published version
Size
1.88 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Chen, Xun
Gentili, Matteo
Hacohen, Nir
Regev, Aviv
Date Issued
December 2021
Journal
Nature Communications
Publisher
Springer Science and Business Media LLC
Citation
Chen, Xun et al. "A cell-free nanobody engineering platform rapidly generates SARS-CoV-2 neutralizing nanobodies." Nature Communications (December 2021): 5506. © 2021 The Author(s)
Version
Final published version
Abstract
Antibody engineering technologies face increasing demands for speed, reliability and scale. We develop CeVICA, a cell-free nanobody engineering platform that uses ribosome display for in vitro selection of nanobodies from a library of 10¹¹ randomized sequences. We apply CeVICA to engineer nanobodies against the Receptor Binding Domain (RBD) of SARS-CoV-2 spike protein and identify >800 binder families using a computational pipeline based on CDR-directed clustering. Among 38 experimentally-tested families, 30 are true RBD binders and 11 inhibit SARS-CoV-2 pseudotyped virus infection. Affinity maturation and multivalency engineering increase nanobody binding affinity and yield a virus neutralizer with picomolar IC50. Furthermore, the capability of CeVICA for comprehensive binder prediction allows us to validate the fitness of our nanobody library. CeVICA offers an integrated solution for rapid generation of divergent synthetic nanobodies with tunable affinities in vitro and may serve as the basis for automated and highly parallel nanobody engineering.
MIT Department
Klarman Cell Observatory (Broad Institute)
Massachusetts Institute of Technology. Department of Biology
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/s41467-021-25777-z
