Author: Haas, Cyrus M.; Francino-Urdaniz, Irene M.; Steiner, Paul J.; Whitehead, Timothy A.
Title: Identification of SARS-CoV-2 S RBD escape mutants using yeast screening and deep mutational scanning Cord-id: t7cyg8ji Document date: 2021_9_22
ID: t7cyg8ji
Snippet: Here we describe a protocol to identify escape mutants on the 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 ten antibodies can be assessed in parallel. This approach has been shown to determine escape mutants that are consistent with more laborious SARS-CoV
Document: Here we describe a protocol to identify escape mutants on the 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 ten antibodies can be assessed in parallel. This approach has been shown to determine escape mutants that are consistent with more laborious SARS-CoV-2 pseudo neutralization assays.
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