Yeast Display and Selections

Abstract

In yeast surface display, yeast cells are exploited to express a protein of interest on their surface, thereby linking it to the encoding DNA within the cell. This display system has become a widely used platform for protein engineering in the past decade, as it confers eukaryotic expression important for the correct assembly of many proteins. In addition, flow cytometry allows for quantitative characterization of binding kinetics and rapid quantitative library screening. In this chapter, we present detailed protocols for yeast surface display and the isolation of naïve binders from a nonimmune scFv library using a combination of magnetic bead selections and fluorescence activated cell sorting. In addition, we describe kinetic binding and thermal stability characterization of proteins expressed via yeast surface display.