1. Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity Nanobody
- Author
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Pedro Chana-Cuevas, German Rehren, Paola Krall, Alberto A. Amarilla, Natalia Lopez-Gonzalez del Rey, Naphak Modhiran, Ananda Müller, Guillermo E. Valenzuela Nieto, Yorka Cheuquemilla, Juan Pablo Toledo, Javier Blesa, Benjamin Uberti, David Schwefel, Ronald Jara, Anne Berking, Teresa Pinto, Constanza Salinas-Rebolledo, Camila Deride, Alexei Cuevas, Daniel Maturana, Daniel Watterson, Luis Ángel Fernández, Héctor Mancilla, Yago Margolles, Pamela Ehrenfeld, Sebastián González-Moraga, Alejandro Rojas-Fernandez, Zaray Miranda-Chacon, Johanna Himelreichs, Alexander A. Khromykh, and Andreas Langer
- Subjects
Male ,Cell biology ,Molecular biology ,Science ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Green Fluorescent Proteins ,Immunology ,Antibody Affinity ,Antibodies, Viral ,Transfection ,Biochemistry ,Article ,Neutralization ,Recombinant antibodies ,Neutralization Tests ,Peptide Library ,Escherichia coli ,Ic50 values ,Animals ,Humans ,Peptide library ,Differential centrifugation ,Multidisciplinary ,SARS-CoV-2 ,Chemistry ,Wild type ,COVID-19 ,Spike Protein ,Single-Domain Antibodies ,Antibodies, Neutralizing ,Surface display ,Virology ,Spike Glycoprotein, Coronavirus ,Medicine ,Immunization ,Angiotensin-Converting Enzyme 2 ,Camelids, New World ,HeLa Cells ,Protein Binding ,Biotechnology ,Healthcare system - Abstract
Despite unprecedented global efforts to rapidly develop SARS-CoV-2 treatments, in order to reduce the burden placed on health systems, the situation remains critical. Effective diagnosis, treatment, and prophylactic measures are urgently required to meet global demand: recombinant antibodies fulfill these requirements and have marked clinical potential. Here, we describe the fast-tracked development of an alpaca Nanobody specific for the receptor-binding-domain (RBD) of the SARS-CoV-2 Spike protein with therapeutic potential applicability. We present a rapid method for nanobody isolation that includes an optimized immunization regimen coupled with VHH library E. coli surface display, which allows single-step selection of high-affinity nanobodies using a simple density gradient centrifugation of the bacterial library. The selected single and monomeric Nanobody, W25, binds to the SARS-CoV-2 S RBD with sub-nanomolar affinity and efficiently competes with ACE-2 receptor binding. Furthermore, W25 potently neutralizes SARS-CoV-2 wild type and the D614G variant with IC50 values in the nanomolar range, demonstrating its potential as antiviral agent.
- Published
- 2020
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