Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display

Abstract Small antibody mimetics that contain high-affinity target-binding peptides can be lower cost alternatives to monoclonal antibodies (mAbs). We have recently developed a method to create small antibody mimetics called FLuctuation-regulated Affinity Proteins (FLAPs), which consist of a small p...

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Autores principales:	Kyra See, Tetsuya Kadonosono, Kotaro Miyamoto, Takuya Tsubaki, Yumi Ota, Marina Katsumi, Sumoe Ryo, Kazuki Aida, Misa Minegishi, Tatsuhiro Isozaki, Takahiro Kuchimaru, Shinae Kizaka-Kondoh
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4a3de7440c4544338c3384a1404a0c94
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spelling	oai:doaj.org-article:4a3de7440c4544338c3384a1404a0c942021-11-14T12:18:29ZAntibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display10.1038/s41598-021-01603-w2045-2322https://doaj.org/article/4a3de7440c4544338c3384a1404a0c942021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01603-whttps://doaj.org/toc/2045-2322Abstract Small antibody mimetics that contain high-affinity target-binding peptides can be lower cost alternatives to monoclonal antibodies (mAbs). We have recently developed a method to create small antibody mimetics called FLuctuation-regulated Affinity Proteins (FLAPs), which consist of a small protein scaffold with a structurally immobilized target-binding peptide. In this study, to further develop this method, we established a novel screening system for FLAPs called monoclonal antibody-guided peptide identification and engineering (MAGPIE), in which a mAb guides selection in two manners. First, antibody-guided design allows construction of a peptide library that is relatively small in size, but sufficient to identify high-affinity binders in a single selection round. Second, in antibody-guided screening, the fluorescently labeled mAb is used to select mammalian cells that display FLAP candidates with high affinity for the target using fluorescence-activated cell sorting. We demonstrate the reliability and efficacy of MAGPIE using daclizumab, a mAb against human interleukin-2 receptor alpha chain (CD25). Three FLAPs identified by MAGPIE bound CD25 with dissociation constants of approximately 30 nM as measured by biolayer interferometry without undergoing affinity maturation. MAGPIE can be broadly adapted to any mAb to develop small antibody mimetics.Kyra SeeTetsuya KadonosonoKotaro MiyamotoTakuya TsubakiYumi OtaMarina KatsumiSumoe RyoKazuki AidaMisa MinegishiTatsuhiro IsozakiTakahiro KuchimaruShinae Kizaka-KondohNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kyra See Tetsuya Kadonosono Kotaro Miyamoto Takuya Tsubaki Yumi Ota Marina Katsumi Sumoe Ryo Kazuki Aida Misa Minegishi Tatsuhiro Isozaki Takahiro Kuchimaru Shinae Kizaka-Kondoh Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display
description	Abstract Small antibody mimetics that contain high-affinity target-binding peptides can be lower cost alternatives to monoclonal antibodies (mAbs). We have recently developed a method to create small antibody mimetics called FLuctuation-regulated Affinity Proteins (FLAPs), which consist of a small protein scaffold with a structurally immobilized target-binding peptide. In this study, to further develop this method, we established a novel screening system for FLAPs called monoclonal antibody-guided peptide identification and engineering (MAGPIE), in which a mAb guides selection in two manners. First, antibody-guided design allows construction of a peptide library that is relatively small in size, but sufficient to identify high-affinity binders in a single selection round. Second, in antibody-guided screening, the fluorescently labeled mAb is used to select mammalian cells that display FLAP candidates with high affinity for the target using fluorescence-activated cell sorting. We demonstrate the reliability and efficacy of MAGPIE using daclizumab, a mAb against human interleukin-2 receptor alpha chain (CD25). Three FLAPs identified by MAGPIE bound CD25 with dissociation constants of approximately 30 nM as measured by biolayer interferometry without undergoing affinity maturation. MAGPIE can be broadly adapted to any mAb to develop small antibody mimetics.
format	article
author	Kyra See Tetsuya Kadonosono Kotaro Miyamoto Takuya Tsubaki Yumi Ota Marina Katsumi Sumoe Ryo Kazuki Aida Misa Minegishi Tatsuhiro Isozaki Takahiro Kuchimaru Shinae Kizaka-Kondoh
author_facet	Kyra See Tetsuya Kadonosono Kotaro Miyamoto Takuya Tsubaki Yumi Ota Marina Katsumi Sumoe Ryo Kazuki Aida Misa Minegishi Tatsuhiro Isozaki Takahiro Kuchimaru Shinae Kizaka-Kondoh
author_sort	Kyra See
title	Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display
title_short	Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display
title_full	Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display
title_fullStr	Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display
title_full_unstemmed	Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display
title_sort	antibody-guided design and identification of cd25-binding small antibody mimetics using mammalian cell surface display
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4a3de7440c4544338c3384a1404a0c94
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Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display

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