Protocol for engineering binding domains to recognize ligand-bound receptors by using yeast surface display.
STAR Protoc
; 5(4): 103339, 2024 Sep 24.
Article
in En
| MEDLINE
| ID: mdl-39321026
ABSTRACT
Yeast surface display is a versatile protein engineering technology, enabling precise control of the applied selection pressure. We present a yeast-surface-display-based protocol for the enrichment of binders specifically recognizing ligand-bound receptors. We describe steps for magnetic bead selections, random mutagenesis, and flow cytometric sorting, followed by library sequencing and detailed analysis of enriched clones. While this approach is exemplified with rcSso7d-based libraries and epidermal growth factor (EGF)-epidermal growth factor receptor (EGFR) complexes, it can also be adapted to other binder scaffolds and ligand-receptor systems. For complete details on the use and execution of this protocol, please refer to Dobersberger et al.1.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
STAR Protoc
Year:
2024
Document type:
Article
Affiliation country:
Austria
Country of publication:
United States