Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
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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; Karl, Vivien; Kim, Changil; Urgard, Egon; Smith, Natalie L; Astorga-Wells, Juan; Ekström, Simon; Coquet, Jonathan M; McInerney, Gerald M; Murrell, Ben.
In: Science Advances, Vol. 8, No. 12, eabm0220, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Multivariate mining of an alpaca immune repertoire identifies potent cross-neutralizing SARS-CoV-2 nanobodies
AU - Hanke, Leo
AU - Sheward, Daniel J
AU - Pankow, Alec
AU - Vidakovics, Laura Perez
AU - Karl, Vivien
AU - Kim, Changil
AU - Urgard, Egon
AU - Smith, Natalie L
AU - Astorga-Wells, Juan
AU - Ekström, Simon
AU - Coquet, Jonathan M
AU - McInerney, Gerald M
AU - Murrell, Ben
PY - 2022
Y1 - 2022
N2 - 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.
AB - 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.
KW - Animals
KW - Antibodies, Monoclonal/chemistry
KW - Antibodies, Viral
KW - COVID-19
KW - Camelids, New World/metabolism
KW - Humans
KW - Membrane Glycoproteins
KW - Neutralization Tests
KW - SARS-CoV-2
KW - Single-Domain Antibodies
KW - Spike Glycoprotein, Coronavirus
KW - Viral Envelope Proteins/metabolism
U2 - 10.1126/sciadv.abm0220
DO - 10.1126/sciadv.abm0220
M3 - Journal article
C2 - 35333580
VL - 8
JO - Science advances
JF - Science advances
SN - 2375-2548
IS - 12
M1 - eabm0220
ER -
ID: 356967727