Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody.
Angiotensin-Converting Enzyme 2
/ immunology
Animals
Antibodies, Neutralizing
/ immunology
Antibodies, Viral
/ immunology
Antibody Affinity
/ genetics
COVID-19
/ immunology
Camelids, New World
/ immunology
Escherichia coli
/ genetics
Green Fluorescent Proteins
/ genetics
HeLa Cells
Humans
Immunization
Male
Neutralization Tests
Peptide Library
Protein Binding
/ genetics
SARS-CoV-2
/ chemistry
Single-Domain Antibodies
/ immunology
Spike Glycoprotein, Coronavirus
/ genetics
Transfection
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
08 02 2021
08 02 2021
Historique:
received:
19
10
2020
accepted:
25
01
2021
entrez:
9
2
2021
pubmed:
10
2
2021
medline:
20
2
2021
Statut:
epublish
Résumé
Despite unprecedented global efforts to rapidly develop SARS-CoV-2 treatments, in order to reduce the burden placed on health systems, the situation remains critical. Effective diagnosis, treatment, and prophylactic measures are urgently required to meet global demand: recombinant antibodies fulfill these requirements and have marked clinical potential. Here, we describe the fast-tracked development of an alpaca Nanobody specific for the receptor-binding-domain (RBD) of the SARS-CoV-2 Spike protein with potential therapeutic applicability. We present a rapid method for nanobody isolation that includes an optimized immunization regimen coupled with VHH library E. coli surface display, which allows single-step selection of Nanobodies using a simple density gradient centrifugation of the bacterial library. The selected single and monomeric Nanobody, W25, binds to the SARS-CoV-2 S RBD with sub-nanomolar affinity and efficiently competes with ACE-2 receptor binding. Furthermore, W25 potently neutralizes SARS-CoV-2 wild type and the D614G variant with IC50 values in the nanomolar range, demonstrating its potential as antiviral agent.
Identifiants
pubmed: 33558635
doi: 10.1038/s41598-021-82833-w
pii: 10.1038/s41598-021-82833-w
pmc: PMC7870875
doi:
Substances chimiques
Antibodies, Neutralizing
0
Antibodies, Viral
0
Peptide Library
0
Single-Domain Antibodies
0
Spike Glycoprotein, Coronavirus
0
spike protein, SARS-CoV-2
0
Green Fluorescent Proteins
147336-22-9
ACE2 protein, human
EC 3.4.17.23
Angiotensin-Converting Enzyme 2
EC 3.4.17.23
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
3318Subventions
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONDEF
ID : ID17I10037
Organisme : FONIS EU-LAC
ID : T010047
Organisme : FONIS EU-LAC
ID : T010047
Organisme : FONIS EU-LAC
ID : T010047
Organisme : FONIS EU-LAC
ID : T010047
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FIC Los Rios
ID : FICR16-19 and FICR19-20
Organisme : FONDECYT No.
ID : 11150532
Organisme : FONDECYT No.
ID : 11150532
Organisme : ISCIII Miguel Servet Program
ID : CP19/00200
Organisme : ISCIII Miguel Servet Program
ID : CP19/00200
Organisme : Agencia Española de Investigación AEI/MICIU/FEDER, EU
ID : Grants BIO2017-89081-R
Organisme : Agencia Española de Investigación AEI/MICIU/FEDER, EU
ID : Grants BIO2017-89081-R
Organisme : Ministerio de Ciencia e Innovación from Spain
ID : CSIC PIE-RDL-COVID-19
Organisme : PAI-CONICYT
ID : 79150075
Organisme : FONDEQUIP EQM
ID : 180037
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