Optimized single-cell gates for yeast display screening

被引:0
|
作者
Pan, Xiaoli [1 ,2 ,3 ]
de Souza, Matheus O. [1 ,2 ,3 ]
Figueiras, Francisco M. [1 ,2 ]
Huang, Aric [3 ]
Banach, Bailey B. [4 ]
Wolfe, Jacy R. [3 ]
Pirhanov, Azady [1 ,2 ]
Madan, Bharat [1 ,2 ,3 ]
Dekosky, Brandon J. [1 ,2 ,3 ,4 ]
机构
[1] Ragon Inst Mass Gen MIT & Harvard, 600 Main St, Cambridge, MA 02139 USA
[2] MIT, Dept Chem Engn, 77 Massachusetts Ave,Room 66-350, Cambridge, MA 02139 USA
[3] Univ Kansas, Dept Pharmaceut Chem, 2093 Constant Ave, Lawrence, KS 66045 USA
[4] Univ Kansas, Bioengn Grad Program, 1530 W 15th St, Lawrence, KS 66045 USA
基金
美国国家卫生研究院;
关键词
Yeast surface display; flow cytometry; single cells; monoclonal antibodies; SURFACE DISPLAY; HUMAN-ANTIBODIES; FLOW-CYTOMETRY;
D O I
10.1093/protein/gzae018
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页数:6
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