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Methods Mol Biol. 2015;1319:3-36. doi: 10.1007/978-1-4939-2748-7_1.

Protein Engineering and Selection Using Yeast Surface Display.

Author information

1
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Building 76-261, 500 Main Street, Cambridge, MA, 02139, USA.

Abstract

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.

PMID:
26060067
DOI:
10.1007/978-1-4939-2748-7_1
[Indexed for MEDLINE]

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