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A cell-free nanobody engineering platform rapidly generates SARS-CoV-2 neutralizing nanobodies
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A cell-free nanobody engineering platform rapidly generates SARS-CoV-2 neutralizing nanobodies
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Journal Article
A cell-free nanobody engineering platform rapidly generates SARS-CoV-2 neutralizing nanobodies
2021
Overview
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 10
11
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.
Faster, higher throughput antibody engineering methods are needed. Here the authors present CeVICA, a cell-free nanobody engineering platform using ribosome display and computational clustering analysis for in vitro selection; they use this to develop nanobodies against the RBD of SARS-CoV-2 spike protein.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 13/31
/ 49/23
/ 49/40
/ 82/16
/ 82/80
/ 82/83
/ 9/10
/ Affinity
/ Antibodies, Neutralizing - immunology
/ Binders
/ Binding
/ COVID-19
/ Humanities and Social Sciences
/ Humans
/ Proteins
/ Science
/ Severe acute respiratory syndrome coronavirus 2
/ Single-Domain Antibodies - chemistry
/ Single-Domain Antibodies - genetics
/ Single-Domain Antibodies - pharmacology
/ Spike Glycoprotein, Coronavirus
/ Viruses
