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Nat Chem Biol. 2016 Feb;12(2):76-81. doi: 10.1038/nchembio.1978. Epub 2015 Dec 7.

High-throughput analysis and protein engineering using microcapillary arrays.

Author information

1
Department of Bioengineering, Stanford University, Stanford, California, USA.
2
Department of Chemical Engineering, Stanford University, Stanford, California, USA.
3
Department of Biochemistry, Stanford University, Stanford, California, USA.
4
Department of Chemistry, Stanford University, Stanford, California, USA.
5
Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, California, USA.
6
Stanford Photonics Research Center, Stanford University, Stanford, California, USA.

Abstract

We describe a multipurpose technology platform, termed μSCALE (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. μSCALE 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 μSCALE, 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 μSCALE platform.

PMID:
26641932
DOI:
10.1038/nchembio.1978
[Indexed for MEDLINE]

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