Identification of SARS-CoV-2 S RBD escape mutants using yeast screening and deep mutational scanning

STAR Protocols

Irene M. Francino-Urdaniz¹  Cyrus M. Haas²  Timothy A. Whitehead²  Paul J. Steiner² 

[1] Corresponding author;Department of Chemical and Biological Engineering, University of Colorado, Boulder, Boulder, CO 80303, USA;

关键词: Sequence analysis;    Sequencing;    Immunology;    Microbiology;    Molecular Biology;    Antibody;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown