tetano
Editor, Senior Moderator
Sci Adv
. 2022 Mar 25;8(12):eabm0220.
doi: 10.1126/sciadv.abm0220. Epub 2022 Mar 25.
Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
Leo Hanke[SUP] 1 [/SUP], Daniel J Sheward[SUP] 1 2 [/SUP], Alec Pankow[SUP] 1 [/SUP], Laura Perez Vidakovics[SUP] 1 [/SUP], Vivien Karl[SUP] 1 [/SUP], Changil Kim[SUP] 1 [/SUP], Egon Urgard[SUP] 1 [/SUP], Natalie L Smith[SUP] 1 [/SUP], Juan Astorga-Wells[SUP] 3 [/SUP], Simon Ekström[SUP] 4 [/SUP], Jonathan M Coquet[SUP] 1 [/SUP], Gerald M McInerney[SUP] 1 [/SUP], Ben Murrell[SUP] 1 [/SUP]
Affiliations
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 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.
. 2022 Mar 25;8(12):eabm0220.
doi: 10.1126/sciadv.abm0220. Epub 2022 Mar 25.
Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
Leo Hanke[SUP] 1 [/SUP], Daniel J Sheward[SUP] 1 2 [/SUP], Alec Pankow[SUP] 1 [/SUP], Laura Perez Vidakovics[SUP] 1 [/SUP], Vivien Karl[SUP] 1 [/SUP], Changil Kim[SUP] 1 [/SUP], Egon Urgard[SUP] 1 [/SUP], Natalie L Smith[SUP] 1 [/SUP], Juan Astorga-Wells[SUP] 3 [/SUP], Simon Ekström[SUP] 4 [/SUP], Jonathan M Coquet[SUP] 1 [/SUP], Gerald M McInerney[SUP] 1 [/SUP], Ben Murrell[SUP] 1 [/SUP]
Affiliations
- PMID: 35333580
- DOI: 10.1126/sciadv.abm0220
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 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.