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Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies

Conventional approaches to isolate and characterize nanobodies are laborious. We combine phage display, multivariate enrichment, next-generation sequencing, and a streamlined screening strategy to identify numerous anti–severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nanobodies. We char...

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Detalles Bibliográficos
Autores principales: Hanke, Leo, Sheward, Daniel J., Pankow, Alec, Vidakovics, Laura Perez, Karl, Vivien, Kim, Changil, Urgard, Egon, Smith, Natalie L., Astorga-Wells, Juan, Ekström, Simon, Coquet, Jonathan M., McInerney, Gerald M., Murrell, Ben
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8956255/
https://www.ncbi.nlm.nih.gov/pubmed/35333580
http://dx.doi.org/10.1126/sciadv.abm0220
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author Hanke, Leo
Sheward, Daniel J.
Pankow, Alec
Vidakovics, Laura Perez
Karl, Vivien
Kim, Changil
Urgard, Egon
Smith, Natalie L.
Astorga-Wells, Juan
Ekström, Simon
Coquet, Jonathan M.
McInerney, Gerald M.
Murrell, Ben
author_facet Hanke, Leo
Sheward, Daniel J.
Pankow, Alec
Vidakovics, Laura Perez
Karl, Vivien
Kim, Changil
Urgard, Egon
Smith, Natalie L.
Astorga-Wells, Juan
Ekström, Simon
Coquet, Jonathan M.
McInerney, Gerald M.
Murrell, Ben
author_sort Hanke, Leo
collection PubMed
description Conventional approaches to isolate and characterize nanobodies are laborious. We combine phage display, multivariate enrichment, next-generation sequencing, and a streamlined screening strategy to identify numerous anti–severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nanobodies. We characterize their potency and specificity using neutralization assays and hydrogen/deuterium exchange mass spectrometry (HDX-MS). The most potent nanobodies bind to the receptor binding motif of the receptor binding domain (RBD), and we identify two exceptionally potent members of this category (with monomeric half-maximal inhibitory concentrations around 13 and 16 ng/ml). Other nanobodies bind to a more conserved epitope on the side of the RBD and are able to potently neutralize the SARS-CoV-2 founder virus (42 ng/ml), the Beta variant (B.1.351/501Y.V2) (35 ng/ml), and also cross-neutralize the more distantly related SARS-CoV-1 (0.46 μg/ml). The approach presented here is well suited for the screening of phage libraries to identify functional nanobodies for various biomedical and biochemical applications.
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spelling pubmed-89562552022-04-04 Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies Hanke, Leo Sheward, Daniel J. Pankow, Alec Vidakovics, Laura Perez Karl, Vivien Kim, Changil Urgard, Egon Smith, Natalie L. Astorga-Wells, Juan Ekström, Simon Coquet, Jonathan M. McInerney, Gerald M. Murrell, Ben Sci Adv Biomedicine and Life Sciences Conventional approaches to isolate and characterize nanobodies are laborious. We combine phage display, multivariate enrichment, next-generation sequencing, and a streamlined screening strategy to identify numerous anti–severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nanobodies. We characterize their potency and specificity using neutralization assays and hydrogen/deuterium exchange mass spectrometry (HDX-MS). The most potent nanobodies bind to the receptor binding motif of the receptor binding domain (RBD), and we identify two exceptionally potent members of this category (with monomeric half-maximal inhibitory concentrations around 13 and 16 ng/ml). Other nanobodies bind to a more conserved epitope on the side of the RBD and are able to potently neutralize the SARS-CoV-2 founder virus (42 ng/ml), the Beta variant (B.1.351/501Y.V2) (35 ng/ml), and also cross-neutralize the more distantly related SARS-CoV-1 (0.46 μg/ml). The approach presented here is well suited for the screening of phage libraries to identify functional nanobodies for various biomedical and biochemical applications. American Association for the Advancement of Science 2022-03-25 /pmc/articles/PMC8956255/ /pubmed/35333580 http://dx.doi.org/10.1126/sciadv.abm0220 Text en Copyright © 2022 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). https://creativecommons.org/licenses/by-nc/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (https://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.
spellingShingle Biomedicine and Life Sciences
Hanke, Leo
Sheward, Daniel J.
Pankow, Alec
Vidakovics, Laura Perez
Karl, Vivien
Kim, Changil
Urgard, Egon
Smith, Natalie L.
Astorga-Wells, Juan
Ekström, Simon
Coquet, Jonathan M.
McInerney, Gerald M.
Murrell, Ben
Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
title Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
title_full Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
title_fullStr Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
title_full_unstemmed Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
title_short Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
title_sort multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing sars-cov-2 nanobodies
topic Biomedicine and Life Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8956255/
https://www.ncbi.nlm.nih.gov/pubmed/35333580
http://dx.doi.org/10.1126/sciadv.abm0220
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