可去除染料--N-聚糖多方法深入分析中的缺失环节  

Removable Dyes-The Missing Link for In-Depth N-Glycan Analysis via Multi-Method Approaches

作　　者：Samanta Cajic RenéHennig Valerian Grote Udo Reichl Erdmann Rapp 

机构地区：[1]Max Planck Institute for Dynamics of Complex Technical Systems,Magdeburg 39106,Germany [2]glyXera GmbH,Magdeburg 39120,Germany [3]Otto-von-Guericke University,Chair of Bioprocess Engineering,Magdeburg 39106,Germany

出　　处：《Engineering》2023年第7期132-150,I0006,共20页工程（英文）

基　　金：support from the German Federal Ministry of Education and Research(BMBF)under the project“Die Golgi Glykan Fabrik 2.0”(031A557C for Samanta Cajic and Erdmann Rapp);the European Commission(EC)under the project“HighGlycan”(278535 for RenéHennig and Erdmann Rapp);the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)under the project“The concert of dolicholbased glycosylation:from molecules to disease models”(FOR2509 for Valerian Grote and Erdmann Rapp).

摘　　要：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(xCGELIF)and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOFMS).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.

关 键 词：GLYCOPROTEINS N-GLYCANS Reversible labeling Hydrophilic interaction liquid CHROMATOGRAPHY Capillary gel electrophoresis Mass spectrometry 

分 类 号：R341[医药卫生—基础医学]