Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches
As the roles of glycans in health and disease continue to be unraveled, it is becoming apparent that glycans’ immense complexity cannot be ignored. To fully delineate glycan structures, we developed an integrative approach combining a set of cost-effective, widespread, and easy-to-handle analytical...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-07-01
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| Series: | Engineering |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095809923002126 |
| _version_ | 1797672757743321088 |
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| author | Samanta Cajic René Hennig Valerian Grote Udo Reichl Erdmann Rapp |
| author_facet | Samanta Cajic René Hennig Valerian Grote Udo Reichl Erdmann Rapp |
| author_sort | Samanta Cajic |
| collection | DOAJ |
| description | As the roles of glycans in health and disease continue to be unraveled, it is becoming apparent that glycans’ immense complexity cannot be ignored. To fully delineate glycan structures, we developed an integrative approach combining a set of cost-effective, widespread, and easy-to-handle analytical methods. The key feature of our workflow is the exploitation of a removable fluorescent label—exemplified by 9-fluorenylmethyl chloroformate (Fmoc)—to bridge the gap between diverse glycoanalytical methods, especially multiplexed capillary gel electrophoresis with laser-induced fluorescence detection (xCGE-LIF) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Through the detailed structural analysis of selected, dauntingly complex N-glycans from chicken ovalbumin, horse serum, and bovine transferrin, we illustrate the capabilities of the presented strategy. Moreover, this approach “visualizes” N-glycans that have been difficult to identify thus far—such as the sulfated glycans on human immunoglobulin A—including minute changes in glycan structures, potentially providing useful new targets for biomarker discovery. |
| first_indexed | 2024-03-11T21:34:52Z |
| format | Article |
| id | doaj.art-79d6f955893c4879851bc7d1e77f8b07 |
| institution | Directory Open Access Journal |
| issn | 2095-8099 |
| language | English |
| last_indexed | 2024-03-11T21:34:52Z |
| publishDate | 2023-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Engineering |
| spelling | doaj.art-79d6f955893c4879851bc7d1e77f8b072023-09-27T04:42:47ZengElsevierEngineering2095-80992023-07-0126132150Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method ApproachesSamanta Cajic0René Hennig1Valerian Grote2Udo Reichl3Erdmann Rapp4Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg 39106, Germany; glyXera GmbH, Magdeburg 39120, GermanyMax Planck Institute for Dynamics of Complex Technical Systems, Magdeburg 39106, Germany; glyXera GmbH, Magdeburg 39120, GermanyMax Planck Institute for Dynamics of Complex Technical Systems, Magdeburg 39106, GermanyMax Planck Institute for Dynamics of Complex Technical Systems, Magdeburg 39106, Germany; Otto-von-Guericke University, Chair of Bioprocess Engineering, Magdeburg 39106, GermanyMax Planck Institute for Dynamics of Complex Technical Systems, Magdeburg 39106, Germany; glyXera GmbH, Magdeburg 39120, Germany; Corresponding author.As the roles of glycans in health and disease continue to be unraveled, it is becoming apparent that glycans’ immense complexity cannot be ignored. To fully delineate glycan structures, we developed an integrative approach combining a set of cost-effective, widespread, and easy-to-handle analytical methods. The key feature of our workflow is the exploitation of a removable fluorescent label—exemplified by 9-fluorenylmethyl chloroformate (Fmoc)—to bridge the gap between diverse glycoanalytical methods, especially multiplexed capillary gel electrophoresis with laser-induced fluorescence detection (xCGE-LIF) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Through the detailed structural analysis of selected, dauntingly complex N-glycans from chicken ovalbumin, horse serum, and bovine transferrin, we illustrate the capabilities of the presented strategy. Moreover, this approach “visualizes” N-glycans that have been difficult to identify thus far—such as the sulfated glycans on human immunoglobulin A—including minute changes in glycan structures, potentially providing useful new targets for biomarker discovery.http://www.sciencedirect.com/science/article/pii/S2095809923002126GlycoproteinsN-glycansReversible labelingHydrophilic interaction liquid chromatographyCapillary gel electrophoresisMass spectrometry |
| spellingShingle | Samanta Cajic René Hennig Valerian Grote Udo Reichl Erdmann Rapp Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches Engineering Glycoproteins N-glycans Reversible labeling Hydrophilic interaction liquid chromatography Capillary gel electrophoresis Mass spectrometry |
| title | Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches |
| title_full | Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches |
| title_fullStr | Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches |
| title_full_unstemmed | Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches |
| title_short | Removable Dyes—The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches |
| title_sort | removable dyes the missing link for in depth n glycan analysis via multi method approaches |
| topic | Glycoproteins N-glycans Reversible labeling Hydrophilic interaction liquid chromatography Capillary gel electrophoresis Mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2095809923002126 |
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