Dobersberger Markus, Sumesgutner Delia, Zajc Charlotte U, Traxlmayr Michael W
Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria.
Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria; CD Laboratory for Next Generation CAR T Cells, Vienna 1090, Austria.
STAR Protoc. 2024 Dec 20;5(4):103339. doi: 10.1016/j.xpro.2024.103339. Epub 2024 Sep 24.
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..
酵母表面展示是一种通用的蛋白质工程技术，能够精确控制施加的选择压力。我们提出了一种基于酵母表面展示的方案，用于富集特异性识别配体结合受体的结合物。我们描述了磁珠筛选、随机诱变和流式细胞术分选的步骤，随后进行文库测序和对富集克隆的详细分析。虽然该方法以基于rcSso7d的文库和表皮生长因子（EGF）-表皮生长因子受体（EGFR）复合物为例，但它也可适用于其他结合物支架和配体-受体系统。有关此方案使用和执行的完整详细信息，请参考多贝斯贝格尔等人的文献。
