Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans
Abstract The study of natural glycans is fundamental to modern glycomics; however, the functional analysis of these glycans is limited by their low natural abundance, structural heterogeneity, and the absence of efficient preparative-scale isolation methods. Here, we present a streamlined approach f...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-04-01
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| Series: | Communications Chemistry |
| Online Access: | https://doi.org/10.1038/s42004-025-01499-x |
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| author | Qing Zhang Yi Lasanajak Menxin Yan Golden Chen Xuezheng Song |
| author_facet | Qing Zhang Yi Lasanajak Menxin Yan Golden Chen Xuezheng Song |
| author_sort | Qing Zhang |
| collection | DOAJ |
| description | Abstract The study of natural glycans is fundamental to modern glycomics; however, the functional analysis of these glycans is limited by their low natural abundance, structural heterogeneity, and the absence of efficient preparative-scale isolation methods. Here, we present a streamlined approach for the gram-scale production of complex natural N-glycans in their reducing form by combining oxidative release of natural glycans (ORNG) using household bleach, an innovative cleavable tag chemistry, and optimized multi-dimensional chromatography. Our ORNG process releases N-glycans from kilogram-scale of natural sources containing various glycoproteins, which can be efficiently attached with a 4-aminobenzoate tag, facilitating the selective separation of these glycans from other biomolecules. The tagged glycans can be purified via dry-load HILIC flash chromatography at gram scale followed by reverse-phase HPLC. Treatment with Oxone quantitatively cleaves the tag, regenerating the free reducing N-glycans in high yield. This method provides an accessible, gram-scale source of complex N-glycans, including complex asymmetric structures that are challenging to synthesize through conventional chemoenzymatic approaches. We believe this approach represents a versatile strategy for acquiring complex natural glycans, opening new avenues for the functional exploration of N-glycans in glycoscience. |
| format | Article |
| id | doaj-art-68f8adcc0a164194a5cfd823ec0bd174 |
| institution | OA Journals |
| issn | 2399-3669 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Chemistry |
| spelling | doaj-art-68f8adcc0a164194a5cfd823ec0bd1742025-08-20T01:53:07ZengNature PortfolioCommunications Chemistry2399-36692025-04-018111210.1038/s42004-025-01499-xStreamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycansQing Zhang0Yi Lasanajak1Menxin Yan2Golden Chen3Xuezheng Song4Department of Biochemistry, Emory University School of MedicineEmory Glycomics and Molecular Interactions Core, Emory University School of MedicineNatGlycan LLCNatGlycan LLCDepartment of Biochemistry, Emory University School of MedicineAbstract The study of natural glycans is fundamental to modern glycomics; however, the functional analysis of these glycans is limited by their low natural abundance, structural heterogeneity, and the absence of efficient preparative-scale isolation methods. Here, we present a streamlined approach for the gram-scale production of complex natural N-glycans in their reducing form by combining oxidative release of natural glycans (ORNG) using household bleach, an innovative cleavable tag chemistry, and optimized multi-dimensional chromatography. Our ORNG process releases N-glycans from kilogram-scale of natural sources containing various glycoproteins, which can be efficiently attached with a 4-aminobenzoate tag, facilitating the selective separation of these glycans from other biomolecules. The tagged glycans can be purified via dry-load HILIC flash chromatography at gram scale followed by reverse-phase HPLC. Treatment with Oxone quantitatively cleaves the tag, regenerating the free reducing N-glycans in high yield. This method provides an accessible, gram-scale source of complex N-glycans, including complex asymmetric structures that are challenging to synthesize through conventional chemoenzymatic approaches. We believe this approach represents a versatile strategy for acquiring complex natural glycans, opening new avenues for the functional exploration of N-glycans in glycoscience.https://doi.org/10.1038/s42004-025-01499-x |
| spellingShingle | Qing Zhang Yi Lasanajak Menxin Yan Golden Chen Xuezheng Song Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans Communications Chemistry |
| title | Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans |
| title_full | Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans |
| title_fullStr | Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans |
| title_full_unstemmed | Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans |
| title_short | Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans |
| title_sort | streamlined gram scale natural n glycan production using reversible tagging after oxidative release of natural glycans |
| url | https://doi.org/10.1038/s42004-025-01499-x |
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