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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 LU ; Karl, Vivien ; Kim, Changil ; Urgard, Egon ; Smith, Natalie L. ; Astorga-Wells, Juan and Ekström, Simon LU , et al. (2022) In Science Advances 8(12).
Abstract

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... (More)

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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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Science Advances
volume
8
issue
12
article number
eabm0220
publisher
American Association for the Advancement of Science (AAAS)
external identifiers
  • pmid:35333580
  • scopus:85127251735
ISSN
2375-2548
DOI
10.1126/sciadv.abm0220
language
English
LU publication?
yes
id
778d5e76-a6ef-4b74-8b37-6141f39ea335
date added to LUP
2022-06-07 13:49:45
date last changed
2025-04-18 15:31:56
@article{778d5e76-a6ef-4b74-8b37-6141f39ea335,
  abstract     = {{<p>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.</p>}},
  author       = {{Hanke, Leo and Sheward, Daniel J. and Pankow, Alec and Vidakovics, Laura Perez and Karl, Vivien and Kim, Changil and Urgard, Egon and Smith, Natalie L. and Astorga-Wells, Juan and Ekström, Simon and Coquet, Jonathan M. and McInerney, Gerald M. and Murrell, Ben}},
  issn         = {{2375-2548}},
  language     = {{eng}},
  number       = {{12}},
  publisher    = {{American Association for the Advancement of Science (AAAS)}},
  series       = {{Science Advances}},
  title        = {{Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies}},
  url          = {{http://dx.doi.org/10.1126/sciadv.abm0220}},
  doi          = {{10.1126/sciadv.abm0220}},
  volume       = {{8}},
  year         = {{2022}},
}