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A cell-free antibody engineering platform rapidly generates SARS-CoV-2 neutralizing antibodies

By Xun Chen, Matteo Gentili, Nir Hacohen, Aviv Regev

Posted 30 Oct 2020
bioRxiv DOI: 10.1101/2020.10.29.361287

Antibody engineering technologies face increasing demands for speed, reliability and scale. We developed CeVICA, a cell-free antibody engineering platform that integrates a novel generation method and design for camelid heavy-chain antibody VHH domain-based synthetic libraries, optimized in vitro selection based on ribosome display and a computational pipeline for binder prediction based on CDR-directed clustering. We applied CeVICA to engineer antibodies against the Receptor Binding Domain (RBD) of the SARS-CoV-2 spike proteins and identified >800 predicted binder families. Among 14 experimentally-tested binders, 6 showed inhibition of pseudotyped virus infection. Antibody affinity maturation further increased binding affinity and potency of inhibition. Additionally, the unique capability of CeVICA for efficient and comprehensive binder prediction allowed retrospective validation of the fitness of our synthetic VHH library design and revealed direction for future refinement. CeVICA offers an integrated solution to rapid generation of divergent synthetic antibodies with tunable affinities in vitro and may serve as the basis for automated and highly parallel antibody generation. ### Competing Interest Statement A.R. is a founder and equity holder of Celsius Therapeutics, an equity holder in Immunitas Therapeutics and until August 31, 2020 was an SAB member of Syros Pharmaceuticals, Neogene Therapeutics, Asimov and ThermoFisher Scientific. From August 1, 2020, A.R. is an employee of Genentech. N.H is an equity holder of BioNtech and is an advisor for Related Sciences. X.C. and A.R. are named co-inventors on a patent application related to CeVICA filed by the Broad Institute that is being made available in accordance with COVID-19 technology licensing framework to maximize access to university innovations.

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