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用于酵母展示筛选的优化单细胞门控

Optimized single-cell gates for yeast display screening.

作者信息

Pan Xiaoli, de Souza Matheus O, Figueiras Francisco M, Huang Aric, Banach Bailey B, Wolfe Jacy R, Pirhanov Azady, Madan Bharat, DeKosky Brandon J

机构信息

The Ragon Institute of Mass General, MIT, and Harvard, 600 Main St, Cambridge, MA 02139, USA.

Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Room 66-350, Cambridge, MA 02139, USA.

出版信息

Protein Eng Des Sel. 2025 Jan 10;38. doi: 10.1093/protein/gzae018.

Abstract

Yeast display is a widely used technology in antibody discovery and protein engineering. The cell size of yeast enables fluorescence-activated cell sorting (FACS) to precisely screen gene libraries, including for multi-parameter selection of protein phenotypes. However, yeast cells show a broader size distribution than mammalian cells that complicates single-cell gate determination for FACS. In this report, we analyze several yeast display gating options in detail and present an optimized strategy to select single yeast cells via flow cytometry. These data reveal optimized single-cell gating strategies to support robust and high-efficiency yeast display studies.

摘要

酵母展示是一种在抗体发现和蛋白质工程中广泛应用的技术。酵母的细胞大小使得荧光激活细胞分选(FACS)能够精确筛选基因文库,包括用于蛋白质表型的多参数选择。然而,酵母细胞的大小分布比哺乳动物细胞更广泛,这使得FACS的单细胞门控确定变得复杂。在本报告中,我们详细分析了几种酵母展示门控选项,并提出了一种通过流式细胞术选择单个酵母细胞的优化策略。这些数据揭示了优化的单细胞门控策略,以支持稳健且高效的酵母展示研究。

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