Development of a yeast cell surface display method using the SpyTag/SpyCatcher system

Abstract Yeast cell surface display (YSD) has been used to engineer various proteins, including antibodies. Directed evolution, which subjects a gene to iterative rounds of mutagenesis, selection and amplification, is useful for protein engineering. In vivo continuous mutagenesis, which continuously...

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Autores principales: Kaho Kajiwara, Wataru Aoki, Naoki Koike, Mitsuyoshi Ueda
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Publicado: Nature Portfolio 2021
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Acceso en línea:https://doaj.org/article/3c96bce5a6a04f1d89df7768632d6aad
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spelling oai:doaj.org-article:3c96bce5a6a04f1d89df7768632d6aad2021-12-02T15:49:42ZDevelopment of a yeast cell surface display method using the SpyTag/SpyCatcher system10.1038/s41598-021-90593-w2045-2322https://doaj.org/article/3c96bce5a6a04f1d89df7768632d6aad2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90593-whttps://doaj.org/toc/2045-2322Abstract Yeast cell surface display (YSD) has been used to engineer various proteins, including antibodies. Directed evolution, which subjects a gene to iterative rounds of mutagenesis, selection and amplification, is useful for protein engineering. In vivo continuous mutagenesis, which continuously diversifies target genes in the host cell, is a promising tool for accelerating directed evolution. However, combining in vivo continuous evolution and YSD is difficult because mutations in the gene encoding the anchor proteins may inhibit the display of target proteins on the cell surface. In this study, we have developed a modified YSD method that utilises SpyTag/SpyCatcher-based in vivo protein ligation. A nanobody fused with a SpyTag of 16 amino acids and an anchor protein fused with a SpyCatcher of 113 amino acids are encoded by separate gene cassettes and then assembled via isopeptide bond formation. This system achieved a high display efficiency of more than 90%, no intercellular protein ligation events, and the enrichment of target cells by cell sorting. These results suggested that our system demonstrates comparable performance with conventional YSD methods; therefore, it can be an appropriate platform to be integrated with in vivo continuous evolution.Kaho KajiwaraWataru AokiNaoki KoikeMitsuyoshi UedaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Kaho Kajiwara
Wataru Aoki
Naoki Koike
Mitsuyoshi Ueda
Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
description Abstract Yeast cell surface display (YSD) has been used to engineer various proteins, including antibodies. Directed evolution, which subjects a gene to iterative rounds of mutagenesis, selection and amplification, is useful for protein engineering. In vivo continuous mutagenesis, which continuously diversifies target genes in the host cell, is a promising tool for accelerating directed evolution. However, combining in vivo continuous evolution and YSD is difficult because mutations in the gene encoding the anchor proteins may inhibit the display of target proteins on the cell surface. In this study, we have developed a modified YSD method that utilises SpyTag/SpyCatcher-based in vivo protein ligation. A nanobody fused with a SpyTag of 16 amino acids and an anchor protein fused with a SpyCatcher of 113 amino acids are encoded by separate gene cassettes and then assembled via isopeptide bond formation. This system achieved a high display efficiency of more than 90%, no intercellular protein ligation events, and the enrichment of target cells by cell sorting. These results suggested that our system demonstrates comparable performance with conventional YSD methods; therefore, it can be an appropriate platform to be integrated with in vivo continuous evolution.
format article
author Kaho Kajiwara
Wataru Aoki
Naoki Koike
Mitsuyoshi Ueda
author_facet Kaho Kajiwara
Wataru Aoki
Naoki Koike
Mitsuyoshi Ueda
author_sort Kaho Kajiwara
title Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
title_short Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
title_full Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
title_fullStr Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
title_full_unstemmed Development of a yeast cell surface display method using the SpyTag/SpyCatcher system
title_sort development of a yeast cell surface display method using the spytag/spycatcher system
publisher Nature Portfolio
publishDate 2021
url https://doaj.org/article/3c96bce5a6a04f1d89df7768632d6aad
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AT wataruaoki developmentofayeastcellsurfacedisplaymethodusingthespytagspycatchersystem
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