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Protein Engineering by Yeast Surface Display
Published on: November 29, 2024
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Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
Kaho Kajiwara1, Wataru Aoki1,2, Naoki Koike3
1Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
Scientific Reports
|May 27, 2021
Summary
This study presents a novel yeast cell surface display (YSD) method using SpyTag/SpyCatcher ligation, enabling efficient protein engineering and directed evolution. The system overcomes previous limitations, achieving high display efficiency and compatibility with continuous mutagenesis for accelerated protein development.
Area of Science:
- Biotechnology
- Protein Engineering
- Synthetic Biology
Background:
- Yeast cell surface display (YSD) is a powerful technique for protein engineering, particularly for antibodies.
- Directed evolution accelerates protein engineering through iterative mutagenesis and selection.
- Integrating in vivo continuous mutagenesis with YSD is challenging due to potential disruptions in anchor protein function.
Purpose of the Study:
- To develop a modified YSD system that facilitates integration with in vivo continuous evolution.
- To overcome limitations of conventional YSD methods that hinder the combination with continuous mutagenesis.
- To establish a robust platform for accelerated protein engineering via directed evolution.
Main Methods:
- Developed a modified YSD system utilizing SpyTag/SpyCatcher-based in vivo protein ligation.
- Engineered separate gene cassettes for a nanobody-fused SpyTag and an anchor protein-fused SpyCatcher.
- Assembled proteins via isopeptide bond formation, enabling modular display on yeast cell surfaces.
Main Results:
- Achieved high display efficiency exceeding 90% for target proteins on the yeast cell surface.
- Demonstrated no intercellular protein ligation events, ensuring system specificity.
- Successfully enriched target cells using cell sorting, validating the system's efficacy.
Conclusions:
- The developed SpyTag/SpyCatcher-based YSD system is a viable platform for protein engineering.
- This method overcomes previous challenges, allowing seamless integration with in vivo continuous evolution.
- The system offers comparable performance to conventional YSD, paving the way for accelerated protein development.
