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Protein Engineering and Selection Using Yeast Surface Display

  • Protocol
Yeast Surface Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1319))

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.

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Acknowledgments

We gratefully acknowledge all members of the Wittrup group for helpful discussions and critical reading of this chapter. Furthermore, we thank the Koch Institute Swanson Biotechnology Center for technical support, specifically the Flow Cytometry and Biopolymers Core Facilities for FACS experiments and DNA sequencing, respectively. The financial contributions from the Swiss National Science Foundation (SNSF Fellowship for Advanced Researchers PA00P3 139659 to A.A.), the NIGMS/MIT Biotechnology Training Program (T.F.C. and N.J.Y.), the ICBP (1 U54 CA112967 to S.D.P.), and the Austrian Science Fund (Schrödinger Fellowship J3496-N28 to M.W.T) are gratefully acknowledged.

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  1. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Building 76-261, 500 Main Street, Cambridge, MA, 02139, USA

    Alessandro Angelini, Tiffany F. Chen, Seymour de Picciotto, Nicole J. Yang, Alice Tzeng, Michael S. Santos, James A. Van Deventer, Michael W. Traxlmayr & K. Dane Wittrup

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

    Alessandro Angelini, Nicole J. Yang, Michael S. Santos, James A. Van Deventer, Michael W. Traxlmayr & K. Dane Wittrup

  3. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Tiffany F. Chen, Seymour de Picciotto, Alice Tzeng & K. Dane Wittrup

Authors
  1. Alessandro Angelini
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  2. Tiffany F. Chen
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  3. Seymour de Picciotto
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  4. Nicole J. Yang
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  5. Alice Tzeng
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  6. Michael S. Santos
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  7. James A. Van Deventer
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  8. Michael W. Traxlmayr
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  9. K. Dane Wittrup
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Corresponding author

Correspondence to K. Dane Wittrup .

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Editors and Affiliations

  1. UCSF, San Francisco, California, USA

    Bin Liu

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© 2015 Springer Science+Business Media New York

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Angelini, A. et al. (2015). Protein Engineering and Selection Using Yeast Surface Display. In: Liu, B. (eds) Yeast Surface Display. Methods in Molecular Biology, vol 1319. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2748-7_1

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  • DOI: https://doi.org/10.1007/978-1-4939-2748-7_1

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2747-0

  • Online ISBN: 978-1-4939-2748-7

  • eBook Packages: Springer Protocols

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