担当区分：筆頭著者   記述言語：英語   掲載種別：研究論文（学術雑誌）   出版者・発行元：Public Library of Science (PLoS)  

Glycan structures hold promise as biomarkers for the early detection of diseases, owing to their sensitive reflection of cellular states. However, glycan analysis remains complex and time-consuming, and the use of hazardous chemicals in traditional hydrazinolysis methods presents a significant barrier to broader application and cross-disciplinary research. To enable efficient glycan biomarker discovery, we aimed to develop a simple and accurate N -glycan analysis method capable of high-throughput sample processing. A key feature of this study is the use of pyridylamination, a fluorescent labeling technique that enables high isomer separation efficiency in reversed-phase LC/MS. After comparing various methods for N -glycan release and purification, we identified Rapid PNGase F (New England Biolabs) and BlotGlyco (Sumitomo Bakelite) as optimal for this application. To improve accuracy by reducing artifact formation, the BlotGlyco protocol was modified from the manufacturer’s original instructions. Using this optimized workflow, we successfully analyzed human serum, human urine, and CHO-K1 membrane fractions, demonstrating that distinct glycan structures in each sample type could be effectively separated and quantified. This work supports the broader adoption of glycan analysis across diverse research fields.

DOI： 10.1371/journal.pone.0336565

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担当区分：筆頭著者   記述言語：英語   掲載種別：機関テクニカルレポート，技術報告書，プレプリント等   出版者・発行元：Springer Science and Business Media LLC  

<p>This protocol describes the purification of N-glycans from approximately 50 µg of protein samples, followed by fluorescent labeling with 2-aminopyridine. The procedure utilizes Rapid PNGase F (New England BioLabs) and BlotGlyco (Sumitomo Bakelite). Representative examples include the preparation of human serum, human urine, and CHO-K1 membrane fractions; however, the protocol is also applicable to other types of samples.</p>

DOI： 10.17504/protocols.io.n2bvjez1pgk5/v1

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開催年月日： 2025年11月

記述言語：日本語   会議種別：ポスター発表  

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記述言語：日本語   会議種別：口頭発表（一般）  

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記述言語：日本語   会議種別：ポスター発表  

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