Identification of SARS-CoV-2 S RBD escape mutants using yeast screening and deep mutational scanning.
STAR Protoc
; 2(4): 100869, 2021 12 17.
Article
en En
| MEDLINE
| ID: mdl-34568839
ABSTRACT
Here, we describe a protocol to identify escape mutants on the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) receptor-binding domain (RBD) using a yeast screen combined with deep mutational scanning. Over 90% of all potential single S RBD escape mutants can be identified for monoclonal antibodies that directly compete with angiotensin-converting enzyme 2 for binding. Six to 10 antibodies can be assessed in parallel. This approach has been shown to determine escape mutants that are consistent with more laborious SARS-CoV-2 pseudoneutralization assays. For complete details on the use and execution of this protocol, please refer to Francino-Urdaniz et al. (2021).
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Saccharomyces cerevisiae
/
Análisis Mutacional de ADN
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Glicoproteína de la Espiga del Coronavirus
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Enzima Convertidora de Angiotensina 2
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SARS-CoV-2
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COVID-19
/
Mutación
Tipo de estudio:
Diagnostic_studies
/
Screening_studies
Límite:
Humans
Idioma:
En
Revista:
STAR Protoc
Año:
2021
Tipo del documento:
Article
País de afiliación:
Estados Unidos