A decade of yeast surface display technology: where are we now?

Abstract

Yeast surface display has become an increasingly popular tool for protein engineering and library screening applications. Recent advances have greatly expanded the capability of yeast surface display, and are highlighted by cell-based selections, epitope mapping, cDNA library screening, and cell adhesion engineering. In this review, we discuss the state-of-the-art yeast display methodologies and the rapidly expanding set of applications afforded by this technology.

Figure 1

Mode of Aga2p linkage may affect expression and function of yeast surface displayed target proteins. (A) Expression construct that fuses the protein of interest by its N-terminus to the C-terminus of Aga2p, via a flexible linker. Extracellular secretion is thus directed by the native Aga2p signal peptide. (B) Construct that fuses the protein of interest by its C-terminus to the N-terminus of Aga2p. A signal peptide must therefore be included at the N-terminus of this construct to direct secretion. In both construct formats, Aga2p is bound to the Aga1p subunit, which is in turn covalently linked to cell wall glucans. A similar cell wall linkage is used by most of the alternate anchoring fusions discussed herein. In both constructs, the protein of interest is typically flanked on its N- and C-terminus by epitope tags. The protein of interest can be oligomeric, in which case one subunit is expressed as an Aga2p fusion while others require a signal peptide to direct them to the secretory pathway for assembly.