Protein Engineering and Selection Using Yeast Surface Display
Protein Engineering and Selection Using Yeast Surface Display
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DOI：
10.1007/978-1-4939-2748-7_1
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发表时间：
2015-01-01
期刊：
影响因子：
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通讯作者：
Wittrup, K. Dane
中科院分区：
文献类型：
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作者：
Angelini, Alessandro;Chen, Tiffany F.;Wittrup, K. Dane
酵母表面展示技术是一项强大的工程技术，用于广泛的蛋白质支架。本方案描述了通过磁珠分离和基于流式细胞术的选择，从酵母上显示的大型组合文库中重新分离蛋白质结合物的过程。分离的单克隆的生物物理特性随后被表征，并通过连续的诱变和流式细胞术选择进一步增强所需的特性，从而使蛋白质结合物对感兴趣的靶蛋白具有更高的稳定性、亲和力和特异性。
Yeast surface display is a powerful technology for engineering a broad range of protein scaffolds. This protocol describes the process for de novo isolation of protein binders from large combinatorial libraries displayed on yeast by using magnetic bead separation followed by flow cytometry-based selection. The biophysical properties of isolated single clones are subsequently characterized, and desired properties are further enhanced through successive rounds of mutagenesis and flow cytometry selections, resulting in protein binders with increased stability, affinity, and specificity for target proteins of interest.