Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody

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 antibodie...

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Autores principales:	Guillermo Valenzuela Nieto, Ronald Jara, Daniel Watterson, Naphak Modhiran, Alberto A. Amarilla, Johanna Himelreichs, Alexander A. Khromykh, Constanza Salinas-Rebolledo, Teresa Pinto, Yorka Cheuquemilla, Yago Margolles, Natalia López González del Rey, Zaray Miranda-Chacon, Alexei Cuevas, Anne Berking, Camila Deride, Sebastián González-Moraga, Héctor Mancilla, Daniel Maturana, Andreas Langer, Juan Pablo Toledo, Ananda Müller, Benjamín Uberti, Paola Krall, Pamela Ehrenfeld, Javier Blesa, Pedro Chana-Cuevas, German Rehren, David Schwefel, Luis Ángel Fernandez, Alejandro Rojas-Fernandez
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ee67da82c4154da782d61078ae559f70
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spelling	oai:doaj.org-article:ee67da82c4154da782d61078ae559f702021-12-02T13:30:22ZPotent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody10.1038/s41598-021-82833-w2045-2322https://doaj.org/article/ee67da82c4154da782d61078ae559f702021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82833-whttps://doaj.org/toc/2045-2322Abstract 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 potential therapeutic 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 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.Guillermo Valenzuela NietoRonald JaraDaniel WattersonNaphak ModhiranAlberto A. AmarillaJohanna HimelreichsAlexander A. KhromykhConstanza Salinas-RebolledoTeresa PintoYorka CheuquemillaYago MargollesNatalia López González del ReyZaray Miranda-ChaconAlexei CuevasAnne BerkingCamila DerideSebastián González-MoragaHéctor MancillaDaniel MaturanaAndreas LangerJuan Pablo ToledoAnanda MüllerBenjamín UbertiPaola KrallPamela EhrenfeldJavier BlesaPedro Chana-CuevasGerman RehrenDavid SchwefelLuis Ángel FernandezAlejandro Rojas-FernandezNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Guillermo Valenzuela Nieto Ronald Jara Daniel Watterson Naphak Modhiran Alberto A. Amarilla Johanna Himelreichs Alexander A. Khromykh Constanza Salinas-Rebolledo Teresa Pinto Yorka Cheuquemilla Yago Margolles Natalia López González del Rey Zaray Miranda-Chacon Alexei Cuevas Anne Berking Camila Deride Sebastián González-Moraga Héctor Mancilla Daniel Maturana Andreas Langer Juan Pablo Toledo Ananda Müller Benjamín Uberti Paola Krall Pamela Ehrenfeld Javier Blesa Pedro Chana-Cuevas German Rehren David Schwefel Luis Ángel Fernandez Alejandro Rojas-Fernandez Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
description	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 potential therapeutic 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 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.
format	article
author	Guillermo Valenzuela Nieto Ronald Jara Daniel Watterson Naphak Modhiran Alberto A. Amarilla Johanna Himelreichs Alexander A. Khromykh Constanza Salinas-Rebolledo Teresa Pinto Yorka Cheuquemilla Yago Margolles Natalia López González del Rey Zaray Miranda-Chacon Alexei Cuevas Anne Berking Camila Deride Sebastián González-Moraga Héctor Mancilla Daniel Maturana Andreas Langer Juan Pablo Toledo Ananda Müller Benjamín Uberti Paola Krall Pamela Ehrenfeld Javier Blesa Pedro Chana-Cuevas German Rehren David Schwefel Luis Ángel Fernandez Alejandro Rojas-Fernandez
author_facet	Guillermo Valenzuela Nieto Ronald Jara Daniel Watterson Naphak Modhiran Alberto A. Amarilla Johanna Himelreichs Alexander A. Khromykh Constanza Salinas-Rebolledo Teresa Pinto Yorka Cheuquemilla Yago Margolles Natalia López González del Rey Zaray Miranda-Chacon Alexei Cuevas Anne Berking Camila Deride Sebastián González-Moraga Héctor Mancilla Daniel Maturana Andreas Langer Juan Pablo Toledo Ananda Müller Benjamín Uberti Paola Krall Pamela Ehrenfeld Javier Blesa Pedro Chana-Cuevas German Rehren David Schwefel Luis Ángel Fernandez Alejandro Rojas-Fernandez
author_sort	Guillermo Valenzuela Nieto
title	Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
title_short	Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
title_full	Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
title_fullStr	Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
title_full_unstemmed	Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
title_sort	potent neutralization of clinical isolates of sars-cov-2 d614 and g614 variants by a monomeric, sub-nanomolar affinity nanobody
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/ee67da82c4154da782d61078ae559f70
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Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody

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