Reverse Glycoblotting Allows Rapid-Enrichment Glycoproteomics of Biopharmaceuticals and Disease-Related Biomarkers

Authors

  • Masaki Kurogochi Dr.,

    1. Division of Advanced Life Science, Graduate School of Advanced Life Science, Frontier Research Center for the Post-Genomic Science and Technology, Hokkaido University, N21, W11, Sapporo 001-0021, Japan, Fax: (+81) 11-706-9042
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  • Maho Amano Dr.,

    1. Division of Advanced Life Science, Graduate School of Advanced Life Science, Frontier Research Center for the Post-Genomic Science and Technology, Hokkaido University, N21, W11, Sapporo 001-0021, Japan, Fax: (+81) 11-706-9042
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  • Masataka Fumoto Dr.,

    1. Shionogi & Co. Ltd., Osaka 553–0002, Japan
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  • Akio Takimoto Dr.,

    1. Shionogi & Co. Ltd., Osaka 553–0002, Japan
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  • Hirosato Kondo Dr.,

    1. Shionogi & Co. Ltd., Osaka 553–0002, Japan
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  • Shin-Ichiro Nishimura Prof. Dr.

    1. Division of Advanced Life Science, Graduate School of Advanced Life Science, Frontier Research Center for the Post-Genomic Science and Technology, Hokkaido University, N21, W11, Sapporo 001-0021, Japan, Fax: (+81) 11-706-9042
    2. Drug-Seeds Discovery Research Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo, 062-8517, Japan
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  • This work was partly supported by funding from the Japan Science and Technology Agency (JST) for “Development of system and technology for advanced measurement and analysis (SENTAN)” and for “Innovation COE Program for Future Drug Discovery and Medical Care”. We are grateful to Dr. Y. Shinohara, Dr. Y. Miura, and Dr. H. Nakagawa for valuable suggestions and discussions on mouse serum N-glycan profiling.

Abstract

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Identifizierung von Glycopeptiden: Eine Hochdurchsatz-Glycoproteomikstrategie wurde mithilfe eines neuartigen, reversen Glycoblottings realisiert. Die Methode umfasst die hoch selektive Oxidation der terminalen Sialinsäurereste von Glycopeptiden und deren nachfolgende Anreicherung durch chemische Ligation mit einem Polymerreagens (siehe Schema; Neu5Ac: N-Acetylneuraminsäure, Neu5Gc: N-Glycolylneuraminsäure, grün: N-Acetyl-D-glucosamin, gelb: D-Mannose, blau: D-Galactose).

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