title: Protein Engineering and Selection Using Yeast Surface Display.
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.
authors.affiliation: Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Building 76-261, 500 Main Street, Cambridge, MA, 02139, USA.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Angelini
authors.forename: Tiffany F
authors.initials: TF
authors.lastname: Chen
authors.forename: Seymour
authors.initials: S
authors.lastname: de Picciotto
authors.forename: Nicole J
authors.initials: NJ
authors.lastname: Yang
authors.forename: Alice
authors.initials: A
authors.lastname: Tzeng
authors.forename: Michael S
authors.initials: MS
authors.lastname: Santos
authors.forename: James A
authors.initials: JA
authors.lastname: Van Deventer
authors.forename: Michael W
authors.initials: MW
authors.lastname: Traxlmayr
authors.forename: K Dane
authors.initials: KD
authors.lastname: Wittrup
mesh_terms: D061505:Cell Surface Display Techniques / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D005434:Flow Cytometry; D059346:Magnets; D016296:Mutagenesis; D019151:Peptide Library; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
chemical_list: D019151:Peptide Library; D011506:Proteins