Protein Engineering and Selection Using Yeast Surface Display.

TitleProtein Engineering and Selection Using Yeast Surface Display.
Publication TypeJournal Article
Year of Publication2015
AuthorsAngelini, A, Chen, TF, de Picciotto, S, Yang, NJ, Tzeng, A, Santos, MS, Van Deventer, JA, Traxlmayr, MW, K Wittrup, D
JournalMethods Mol Biol
Date Published2015
KeywordsCell Surface Display Techniques, Combinatorial Chemistry Techniques, Flow Cytometry, Magnets, Mutagenesis, Peptide Library, Protein Binding, Proteins, Saccharomyces cerevisiae

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.

Alternate JournalMethods Mol. Biol.
PubMed ID26060067
Grant List1 U54 CA112967 / CA / NCI NIH HHS / United States