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Classification of Extracellular Vesicles based on Surface Glycan Structures by Spongy-like Separation Media

View ORCID ProfileEisuke Kanao, Shuntaro Wada, View ORCID ProfileHiroshi Nishida, View ORCID ProfileTakuya Kubo, Tetsuya Tanigawa, View ORCID ProfileKoshi Imami, View ORCID ProfileAsako Shimoda, Kaori Umezaki, View ORCID ProfileYoshihiro Sasaki, View ORCID ProfileKazunari Akiyoshi, View ORCID ProfileJun Adachi, View ORCID ProfileKoji Otsuka, View ORCID ProfileYasushi Ishihama
doi: https://doi.org/10.1101/2022.05.10.491426
Eisuke Kanao
aGraduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
bNational Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan
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  • ORCID record for Eisuke Kanao
  • For correspondence: kanao.eisuke.7s@kyoto-u.ac.jp kubo.takuya.6c@kyoto-u.ac.jp
Shuntaro Wada
cDepartment of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Hiroshi Nishida
aGraduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
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  • ORCID record for Hiroshi Nishida
Takuya Kubo
cDepartment of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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  • For correspondence: kanao.eisuke.7s@kyoto-u.ac.jp kubo.takuya.6c@kyoto-u.ac.jp
Tetsuya Tanigawa
cDepartment of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Koshi Imami
aGraduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
dPrecursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
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Asako Shimoda
eDepartment of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Kaori Umezaki
eDepartment of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Yoshihiro Sasaki
eDepartment of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Kazunari Akiyoshi
eDepartment of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Jun Adachi
aGraduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
bNational Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan
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Koji Otsuka
cDepartment of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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Yasushi Ishihama
aGraduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan
bNational Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan
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Abstract

Extracellular vesicles (EVs) are lipid bilayer vesicles that enclose various biomolecules. EVs hold promise as sensitive biomarkers to detect and monitor various diseases. However, they have heterogenous molecular compositions. The compositions of EVs from identical donor cells obtained using the same purification methods may differ, which is a significant obstacle for elucidating objective biological functions. Herein the potential of a novel lectin-based affinity chromatography (LAC) method to classify EVs based on their glycan structures is demonstrated. The proposed method utilizes a spongy-like monolithic polymer (spongy monolith, SPM), which consists of poly(ethylene-co-glycidyl methacrylate) with continuous micropores and allows an efficient in-situ protein reaction with epoxy groups. Two distinct lectins with different specificities, Sambucus sieboldiana agglutinin and concanavalin A, are effectively immobilized on SPM without impacting the binding activity. Moreover, high recovery rates of liposomal nanoparticles as a model of EVs are achieved due to the large flow-through pores (>10 μm) of SPM. Finally, lectin-immobilized SPMs are employed to classify EVs based on the surface glycan structures and demonstrate different subpopulations by proteome profiling.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted May 11, 2022.
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Classification of Extracellular Vesicles based on Surface Glycan Structures by Spongy-like Separation Media
Eisuke Kanao, Shuntaro Wada, Hiroshi Nishida, Takuya Kubo, Tetsuya Tanigawa, Koshi Imami, Asako Shimoda, Kaori Umezaki, Yoshihiro Sasaki, Kazunari Akiyoshi, Jun Adachi, Koji Otsuka, Yasushi Ishihama
bioRxiv 2022.05.10.491426; doi: https://doi.org/10.1101/2022.05.10.491426
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Classification of Extracellular Vesicles based on Surface Glycan Structures by Spongy-like Separation Media
Eisuke Kanao, Shuntaro Wada, Hiroshi Nishida, Takuya Kubo, Tetsuya Tanigawa, Koshi Imami, Asako Shimoda, Kaori Umezaki, Yoshihiro Sasaki, Kazunari Akiyoshi, Jun Adachi, Koji Otsuka, Yasushi Ishihama
bioRxiv 2022.05.10.491426; doi: https://doi.org/10.1101/2022.05.10.491426

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