期刊
NATURE CHEMICAL BIOLOGY
卷 12, 期 2, 页码 76-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.1978
关键词
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资金
- Stanford-Wallace H. Coulter Translational Partnership Award Program
- Siebel Stem Cell Institute
- Thomas and Stacey Siebel Foundation
- Stanford Photonics Research Center
- Hitachi America Faculty Scholar Award
- US National Institutes of Health [GM49243]
- National Science Foundation Graduate Fellowship Program
- Howard Hughes Medical Institute International Student Research Program
- Fannie and John Hertz Foundation Graduate Fellowship
- Stanford Bio-X Fellowship Program
- Stanford Graduate Fellowship Program
- Stanford Dean's Fellowship Program
We describe a multipurpose technology platform, termed mu SCALE (microcapillary single-cell analysis and laser extraction), that enables massively parallel, quantitative biochemical and biophysical measurements on millions of protein variants expressed from yeast or bacteria. mu SCALE spatially segregates single cells within a microcapillary array, enabling repeated imaging, cell growth and protein expression. We performed high-throughput analysis of cells and their protein products using a range of fluorescent assays, including binding-affinity measurements and dynamic enzymatic assays. A precise laser-based extraction method allows rapid recovery of live clones and their genetic material from microcapillaries for further study. With mu SCALE, we discovered a new antibody against a clinical cancer target, evolved a fluorescent protein biosensor and engineered an enzyme to reduce its sensitivity to its inhibitor. These protein analysis and engineering applications each have unique assay requirements and different host organisms, highlighting the flexibility and technical capabilities of the mSCALE platform.
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