Yeast Surface Display

Overview

Yeast surface display physically couples the protein to be engineered to an anchor protein embedded within the yeast cell wall. The eukaryotic secretory pathway of yeast allows a wide variety of proteins to be displayed, including many with complicated folds. Proteins that have been engineered using yeast surface display include scFvs (Feldhaus and Siegel 2004; Chao et al. 2006), Fab antibody fragments (Weaver-Feldhaus et al. 2004), single-chain T-cell receptors (scTCRs) (Shusta et al. 1999), single-chain major histocompatibility complexes (scMHCs) (Esteban and Zhao 2004), human epidermal growth factor (EGF) (Cochran et al. 2006), cytokines (Rao et al. 2003), epidermal growth factor receptor (EGFR) extracellular domains (Kim et al. 2006), fibronectin type III domains (Lipovsek et al. 2007b; Hackel et al. 2008), cystine-knot peptides (Kimura et al. 2009; Silverman et al. 2009), and horseradish peroxidase (Lipovsek et al. 2007a). The ability of this eukaryotic system to display mammalian proteins has also allowed screening of human cDNA libraries (Wadle et al. 2005; Bidlingmaier and Liu 2006). Figure 2.3 illustrates representative yeast surface display platforms.

Yeast Surface Display Platforms

Early work in yeast surface display involved immobilization of antigens for vaccine development and surface display of enzymes for biocatalysts (Schreuder et al. 1993; Schreuder et al. 1996). While most efforts have focused on using Saccharomyces cerevisiae, other yeast such as Pichia pastoris have also been explored (Tanino et al. 2006). Progress in using yeast surface display formats for applications other than protein engineering has recently been reviewed (Kondo and Ueda 2004).

The vast majority of protein engineering efforts with yeast surface display have been performed using the Aga1-Aga2 display format in S. cerevisiae, a technology developed by Wittrup and coworkers over a decade ago (Figure 2.3A) (Boder and

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Displayed protein

Epitope tag

Anchor domain

Yeast cell wall

N, C

Protein termini

0 0

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