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Updated: Apr 11, 2026

Protein Engineering by Yeast Surface Display
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Yeast Display-Based Antibody Affinity Maturation Using Detergent-Solubilized Cell Lysates.

Benjamin J Tillotson1, Jason M Lajoie, Eric V Shusta

  • 1Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 3631 Engineering Hall, 1415 Engineering Drive, Madison, WI, 53706, USA.

Methods in Molecular Biology (Clifton, N.J.)
|June 11, 2015
PubMed
Summary

Researchers developed a new method using yeast display to find antibodies targeting membrane proteins (MPs). This technique uses cell lysates, overcoming the insolubility issues of MPs for antibody discovery and optimization.

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Area of Science:

  • Biochemistry
  • Immunology
  • Molecular Biology

Background:

  • Membrane proteins (MPs) are crucial drug targets, but their insolubility hinders antibody development.
  • Existing in vitro antibody discovery platforms struggle with membrane protein antigens.

Purpose of the Study:

  • To adapt yeast display for identifying antibodies against membrane proteins.
  • To develop a protocol for antibody optimization using detergent-solubilized cell lysates.

Main Methods:

  • Yeast display technology was modified to utilize detergent-solubilized cell lysates.
  • A kinetic screening approach was implemented to select for antibodies with improved affinity.

Main Results:

  • Successfully integrated cell lysates into the yeast display workflow for membrane protein antigen presentation.

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  • Demonstrated the ability to isolate antibodies with enhanced binding kinetics (slowed dissociation) to membrane proteins.

    • Conclusions:

    • Yeast display is a viable platform for antibody discovery against challenging membrane protein targets.
    • The described protocol enables the engineering of high-affinity antibodies for membrane proteins using cell lysates.