A rapid cell-free expression and screening platform for antibody discovery

Andrew C. Hunt, Bastian Vögeli, Ahmed O. Hassan, Laura Guerrero, Weston Kightlinger, Danielle J. Yoesep, Antje Krüger, Madison DeWinter, Michael S. Diamond, Ashty S. Karim and Michael C. Jewett ()
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Andrew C. Hunt: Northwestern University
Bastian Vögeli: Northwestern University
Ahmed O. Hassan: Washington University School of Medicine
Laura Guerrero: Northwestern University
Weston Kightlinger: Northwestern University
Danielle J. Yoesep: Northwestern University
Antje Krüger: Northwestern University
Madison DeWinter: Northwestern University
Michael S. Diamond: Washington University School of Medicine
Ashty S. Karim: Northwestern University
Michael C. Jewett: Northwestern University

Nature Communications, 2023, vol. 14, issue 1, 1-14

Abstract: Abstract Antibody discovery is bottlenecked by the individual expression and evaluation of antigen-specific hits. Here, we address this bottleneck by developing a workflow combining cell-free DNA template generation, cell-free protein synthesis, and binding measurements of antibody fragments in a process that takes hours rather than weeks. We apply this workflow to evaluate 135 previously published antibodies targeting the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), including all 8 antibodies previously granted emergency use authorization for coronavirus disease 2019 (COVID-19), and demonstrate identification of the most potent antibodies. We also evaluate 119 anti-SARS-CoV-2 antibodies from a mouse immunized with the SARS-CoV-2 spike protein and identify neutralizing antibody candidates, including the antibody SC2-3, which binds the SARS-CoV-2 spike protein of all tested variants of concern. We expect that our cell-free workflow will accelerate the discovery and characterization of antibodies for future pandemics and for research, diagnostic, and therapeutic applications more broadly.

Date: 2023
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DOI: 10.1038/s41467-023-38965-w

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