Glycan Sequencing Based on Glycosidase-Assisted Nanopore Sensing
文献类型：期刊论文
| 作者 | Yao, Guangda6,7,8; Xia, Bingqing5,8; Wei, Fangyu4,5; Wang, Jiahong3; Yang, Yuting5,8; Ma, Shengzhou5; Ke, Wenjun5,8; Li, Tiehai4,5; Cheng, Xi2,8; Wen, Liuqing4,5 |
| 刊名 | JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
 |
| 出版日期 | 2025-01-02 |
| 卷号 | 147期号:2页码:1721-1731 |
| ISSN号 | 0002-7863 |
| DOI | 10.1021/jacs.4c12940 |
| 英文摘要 | Nanopores are promising sensors for glycan analysis with the accurate identification of complex glycans laying the foundation for nanopore-based sequencing. However, their applicability toward continuous glycan sequencing has not yet been demonstrated. Here, we present a proof-of-concept of glycan sequencing by combining nanopore technology with glycosidase-hydrolyzing reactions. By continuously monitoring the changes in the characteristic current generated by the translocation of glycan hydrolysis products through a nanopore, the glycan sequence can be accurately identified based on the specificity of glycosidases. With machine learning, we improved the sequencing accuracy to over 98%, allowing for the reliable determination of consecutive building blocks and glycosidic linkages of glycan chains while reducing the need for operator expertise. This approach was validated on real glycan samples, with accuracy calibrated using hydrophilic interaction chromatography-high-performance liquid chromatography (HILIC-HPLC) and mass spectrometry (MS). We achieved the sequencing of ten consecutive units in natural glycan chains, which provided the first evidence for the feasibility of a nanopore-glycosidase-compatible system in glycan sequencing. Compared to traditional methods, this strategy enhances sequencing efficiency by over 5-fold. Additionally, we introduced the concept of 'inverse sequencing', which focuses on electrical signal changes rather than monosaccharide identification. This eliminates the reliance on glycan fingerprint libraries typically required in putative 'forward hydrolysis' strategies. When the challenges in both 'forward and inverse hydrolysis sequencing strategies' are addressed, this approach will pave the way for establishing a glycan sequencing technology at a single-molecule level. |
| WOS关键词 | ALPHA-HEMOLYSIN |
| 资助项目 | Natural Science Foundation of Shanghai Municipality ; Shanghai Municipal Science and Technology Major Project[92478111,92169202] ; Shanghai Municipal Science and Technology Major Project[82341058] ; Natural Science Foundation of China[2019285] ; Natural Science Foundation of China[2021333] ; Natural Science Foundation of China[2022077] ; Fund of Youth Innovation Promotion Association[20ZR1474200] ; National Fund of Shanghai Science and Technology Innovation Action Plan[22QA1411000] ; Shanghai Rising-Star Program[22ZR1474000] ; Natural Science Foundation of Shanghai[LG202101-01-04] ; Natural Science Foundation of Shanghai[LG202102-01-01] ; National Key Laboratory Program of China[CASSHB-QNPD-2023-018] ; Talent Plan of Shanghai Branch, Chinese Academy of Sciences |
| WOS研究方向 | Chemistry |
| 语种 | 英语 |
| WOS记录号 | WOS:001388706900001 |
| 出版者 | AMER CHEMICAL SOC |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/315591]  |
| 专题 | 新药研究国家重点实验室 |
| 通讯作者 | Xia, Bingqing; Wen, Liuqing; Long, Yi-tao; Gao, Zhaobing |
| 作者单位 | 1.Chinese Acad Sci, Zhongshan Inst Drug Discovery, Shanghai Inst Mat Med, Zhongshan 528400, Peoples R China 2.Hangzhou Inst Adv Study, Sch Pharmaceut Sci & Technol, Hangzhou 330106, Peoples R China 3.Nanjing Univ, Sch Chem & Chem Engn, Nanjing 210023, Peoples R China 4.Chinese Acad Sci, Shanghai Inst Mat Med, Carbohydrate Based Drug Res Ctr, Shanghai 201203, Peoples R China 5.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 6.Lingang Lab, Shanghai 200031, Peoples R China 7.Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China 8.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Yao, Guangda,Xia, Bingqing,Wei, Fangyu,et al. Glycan Sequencing Based on Glycosidase-Assisted Nanopore Sensing[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,2025,147(2):1721-1731. |
| APA | Yao, Guangda.,Xia, Bingqing.,Wei, Fangyu.,Wang, Jiahong.,Yang, Yuting.,...&Gao, Zhaobing.(2025).Glycan Sequencing Based on Glycosidase-Assisted Nanopore Sensing.JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,147(2),1721-1731. |
| MLA | Yao, Guangda,et al."Glycan Sequencing Based on Glycosidase-Assisted Nanopore Sensing".JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 147.2(2025):1721-1731. |
入库方式： OAI收割
来源：上海药物研究所
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