Protein Engineering and Selection Using Yeast Surface Display

Autor: Alessandro, Angelini, Tiffany F, Chen, Seymour, de Picciotto, Nicole J, Yang, Alice, Tzeng, Michael S, Santos, James A, Van Deventer, Michael W, Traxlmayr, K Dane, Wittrup
Rok vydání: 2015
Předmět:
Zdroj: Methods in molecular biology (Clifton, N.J.). 1319
ISSN: 1940-6029
Popis: 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.
Databáze: OpenAIRE