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

Research output: Contribution to journalJournal articleResearchpeer-review

  • Leo Hanke
  • Daniel J Sheward
  • Alec Pankow
  • Laura Perez Vidakovics
  • Vivien Karl
  • Changil Kim
  • Urgard, Egon
  • Natalie L Smith
  • Juan Astorga-Wells
  • Simon Ekström
  • Coquet, Jonathan
  • Gerald M McInerney
  • Ben Murrell

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.

Original languageEnglish
Article numbereabm0220
JournalScience Advances
Issue number12
Publication statusPublished - 2022
Externally publishedYes

    Research areas

  • Animals, Antibodies, Monoclonal/chemistry, Antibodies, Viral, COVID-19, Camelids, New World/metabolism, Humans, Membrane Glycoproteins, Neutralization Tests, SARS-CoV-2, Single-Domain Antibodies, Spike Glycoprotein, Coronavirus, Viral Envelope Proteins/metabolism

ID: 356967727