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Yeast Display and Selections

  • Protocol
Antibody Engineering

Part of the book series: Springer Protocols Handbooks ((SPH))

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.

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Authors and Affiliations

  1. Chemical Engineering Department, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Kelly Davis Orcutt & K. Dane Wittrup

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  1. Kelly Davis Orcutt
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  2. K. Dane Wittrup
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Correspondence to K. Dane Wittrup .

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Editors and Affiliations

  1. (Biomedical Engineering) Institut für Zellbiologie und Immunologie, Universität Stuttgart, Allmandring 31, 70569, Stuttgart, Germany

    Roland Kontermann

  2. Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Spielmannstraße 7, 38106, Braunschweig, Germany

    Stefan Dübel

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© 2010 Springer-Verlag

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Orcutt, K.D., Wittrup, K.D. (2010). Yeast Display and Selections. In: Kontermann, R., Dübel, S. (eds) Antibody Engineering. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-01144-3_15

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  • DOI: https://doi.org/10.1007/978-3-642-01144-3_15

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-01143-6

  • Online ISBN: 978-3-642-01144-3

  • eBook Packages: Springer Protocols

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