Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity Nanobody
| Autor: | 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, Andreas Langer |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
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 |
| Zdroj: | Scientific Reports Scientific Reports, Vol 11, Iss 1, Pp 1-14 (2021) |
| DOI: | 10.1101/2020.06.09.137935 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|