Directed evolution of potent neutralizing nanobodies against SARS-CoV-2 using CDR-swapping mutagenesis. Issue 9 (16th September 2021)

• Record Type: Journal Article
• Authors: Zupancic, Jennifer M.; Desai, Alec A.; Schardt, John S.; Pornnoppadol, Ghasidit; Makowski, Emily K.; Smith, Matthew D.; Kennedy, Andrew A.; Garcia de Mattos Barbosa, Mayara; Cascalho, Marilia; Lanigan, Thomas M.; Tai, Andrew W.; Tessier, Peter M.
• Is Part Of: Cell chemical biology. Volume 28:Issue 9(2021)
• Page Start: 1379
• Page End: 1388.e7
• Publication Date: 2021-09-16
• Subjects: COVID-19 -- neutralization -- affinity -- maturation -- nanobody -- yeast -- receptor-binding domain -- RBD -- ACE2 -- spike; Biochemistry -- Periodicals; 572.05
• DOI: 10.1016/j.chembiol.2021.05.019
• Languages: English
• ISSNs: 2451-9456

Abstract

Summary: There is widespread interest in facile methods for generating potent neutralizing antibodies, nanobodies, and other affinity proteins against SARS-CoV-2 and related viruses to address current and future pandemics. While isolating antibodies from animals and humans are proven approaches, these methods are limited to the affinities, specificities, and functional activities of antibodies generated by the immune system. Here we report a surprisingly simple directed evolution method for generating nanobodies with high affinities and neutralization activities against SARS-CoV-2. We demonstrate that complementarity-determining region swapping between low-affinity lead nanobodies, which we discovered unintentionally but find is simple to implement systematically, results in matured nanobodies with unusually large increases in affinity. Importantly, the matured nanobodies potently neutralize both SARS-CoV-2 pseudovirus and live virus, and possess drug-like biophysical properties. We expect that our methods will improve in vitro nanobody discovery and accelerate the generation of potent neutralizing nanobodies against diverse coronaviruses. Graphical abstract: Highlights: Directed evolution of nanobodies that potently neutralize SARS-CoV-2 CDR-swapping mutagenesis facilitates large affinity and activity improvements Nanobody binding to RBD competes with ACE2 and two classes of neutralizing mAbs Neutralizing nanobodies display drug-like biophysical properties Abstract : Zupancic et al. report potent neutralizing nanobodies against SARS-CoV-2. They demonstrate an approach that involves swapping the complementarity-determining regions of low-affinity clones to generate matured nanobodies with large increases in affinity and neutralization activity.