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Affinity and Stability Analysis of Yeast Displayed Proteins

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Yeast Surface Display

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2491))

Abstract

Yeast surface display is a powerful protein engineering technology that is extensively used to improve various properties of proteins, including affinity, specificity, and stability or even to add novel functions (usually ligand binding). Apart from its robustness and versatility as an engineering tool, yeast display offers a further critical advantage: Once the selection campaign is finished, usually resulting in an oligoclonal pool, these enriched protein variants can be analyzed individually on the surface of yeast without the need for any sub-cloning, soluble expression, and purification. Here, we provide detailed protocols for determining both the affinity and the thermal stability of yeast displayed proteins. In addition, we discuss the advantages, challenges, and potential pitfalls associated with affinity and stability analysis using yeast surface display.

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Acknowledgments

This work was supported by the Austrian Science Fund (FWF Project W1224—Doctoral Program on Biomolecular Technology of Proteins—BioToP) and by the Federal Ministry for Digital and Economic Affairs of Austria and the National Foundation for Research, Technology and Development of Austria to the Christian Doppler Research Association (Christian Doppler Laboratory for Next Generation CAR T Cells).

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Authors and Affiliations

  1. Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, Austria

    Charlotte U. Zajc, Magdalena Teufl & Michael W. Traxlmayr

  2. CD Laboratory for Next Generation CAR T Cells, Vienna, Austria

    Charlotte U. Zajc & Magdalena Teufl

Authors
  1. Charlotte U. Zajc
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  2. Magdalena Teufl
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  3. Michael W. Traxlmayr
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Corresponding author

Correspondence to Michael W. Traxlmayr .

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Editors and Affiliations

  1. Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences, Vienna, Austria

    Michael W. Traxlmayr

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Zajc, C.U., Teufl, M., Traxlmayr, M.W. (2022). Affinity and Stability Analysis of Yeast Displayed Proteins. In: Traxlmayr, M.W. (eds) Yeast Surface Display. Methods in Molecular Biology, vol 2491. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2285-8_9

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  • DOI: https://doi.org/10.1007/978-1-0716-2285-8_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2284-1

  • Online ISBN: 978-1-0716-2285-8

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