Specific Analysis of α-2,3-Sialylated N-Glycan Linkage Isomers by Microchip Capillary Electrophoresis–Mass Spectrometry

Cheng, Mengxia; Shu, Hong; Ye, Peng; Feng, Xiaoxiao; Yan, Guoquan; Zhang, Lei; Yao, Jun; Bao, Huimin; Lu, Haojie

doi:10.1021/acs.analchem.1c00064

Cited by 27 publications

(22 citation statements)

References 48 publications

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“…The reaction mixture was incubated at 37 °C for 2 h and added with 80 μL of ACN and 300 μL of 80% (v/v) ACN containing 0.1% (v/v) TFA to terminate the reaction. The solution was then purified by cotton-wool-based HILIC-SPE …”

Section: Experimental Sectionmentioning

confidence: 99%

“…Meanwhile, the interface of MCE-MS avoids sample dilution in sheath flow interfaces which can results in low sensitivity, and solve the problems of the stability and reproducibility of spray in sheathless-flow interfaces. This MCE-MS technique has been applied into the glycosylation study. , To the best of our knowledge, there was no report about specific analysis of α-2,3- and α-2,6-linked sialylated N-glycan isomers using MCE-MS.…”

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confidence: 99%

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Fast Discrimination of Sialylated N-Glycan Linkage Isomers with One-Step Derivatization by Microfluidic Capillary Electrophoresis–Mass Spectrometry

et al. 2022

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Linkage isomers (α-2,3-or α-2,6-linkage) of sialylated N-glycans are involved in the emergence and progression of some diseases, so they are of great significance for diagnosing and monitoring diseases. However, the qualitative and quantitative analysis of sialylated N-glycan linkage isomers remains challenging due to their low abundance and limited isomeric separation techniques. Herein, we developed a novel strategy integrating one-step sialic acid derivatization, positive charge-sensitive separation and highly sensitive detection based on microfluidic capillary electrophoresis−mass spectrometry (MCE-MS) for fast and specific analysis of α-2,3and α-2,6-linked sialylated N-glycan isomers. A kind of easily charged long-chain amino compound was screened first for one-step sialic acid derivatization so that only α-2,3and α-2,6-linked isomers can be quickly and efficiently separated within 10 min by MCE due to the difference in structural conformation, whose separation mechanism was further theoretically supported by molecular dynamic simulation. In addition, different sialylated N-glycans were separated in order according to the number of sialic acids, so that a migration time-based prediction of the number of sialic acids was achieved. Finally, the sialylated N-glycome of human serum was profiled within 10 min and 6 of the 52 detected sialylated N-glycans could be potential diagnostic biomarkers of cervical cancer (CC), whose α-2,3and α-2,6-linked isomers were distinguished by α-2,3Neuraminidase S.

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Section: Experimental Sectionmentioning

confidence: 99%

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confidence: 99%

Fast Discrimination of Sialylated N-Glycan Linkage Isomers with One-Step Derivatization by Microfluidic Capillary Electrophoresis–Mass Spectrometry

et al. 2022

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“…Cheng et al. [73] designed a strategy that couples the linkage‐specific derivatization with a microchip CE‐MS for the specific analysis of α‐2,3‐sialylated glycans. The carboxyl groups of α‐2,6‐ and α‐2,3‐linked sialic acids were selectively derivatized with methylamine and N , N ‐dimethyl ethylenediamine (DMEN), respectively.…”

Section: Derivatization Of Glycansmentioning

confidence: 99%

“…Lageveen-Kammeijer et al [72] have reported a method that involves ethyl esterification of α-2,6-linked sialic acid and amidation of α-2,3 linkage in a simple two-step reaction. The one-pot reaction with a simple clean-up step allowed distinguishing and high-sensitivity characterization of sialic acid isomers from biofluids and tissues by CE-ESI-MS. Cheng et al [73] designed a strategy that couples the linkage-specific derivatization with a microchip CE-MS for the specific analysis of α-2,3-sialylated glycans. The carboxyl groups of α-2,6and α-2,3-linked sialic acids were selectively derivatized with methylamine and N,N-dimethyl ethylenediamine (DMEN), respectively.…”

Section: Derivatization Techniques Prompting Isomeric Identificationmentioning

confidence: 99%

Advances in mass spectrometry‐based glycomics—An update covering the period 2017–2021

et al. 2021

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The wide variety of chemical properties and biological functions found in proteins is attained via post‐translational modifications like glycosylation. Covalently bonded to proteins, glycans play a critical role in cell activity. Complex structures with microheterogeneity, the glycan structures that are associated with proteins are difficult to analyze comprehensively. Recent advances in sample preparation methods, separation techniques, and MS have facilitated the quantitation and structural elucidation of glycans. This review focuses on highlighting advances in MS‐based techniques for glycomic analysis that occurred over the last 5 years (2017–2021) as an update to the previous review on the subject. The topics of discussion will include progress in glycomic workflow such as glycan release, purification, derivatization, and separation as well as the topics of ionization, tandem MS, and separation techniques that can be coupled with MS. Additionally, bioinformatics tools used for the analysis of glycans will be described.

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“…Despite the importance of sialic acid epitopes in several health issues, accurate identification and quantification of sialylated glycans at the isomeric level remain a challenge due to the extremely heterogeneous sialyation and labile sialic acid residues ( Cheng et al, 2021 ). Today, selection/multiple response monitoring (SRM/MRM) coupled with liquid chromatography (LC) is emerging as an ideal technique for glycan quantitation ( Zhou et al, 2015 ; Seo et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Isomer-Specific Monitoring of Sialylated N-Glycans Reveals Association of α2,3-Linked Sialic Acid Epitope With Behcet’s Disease

Seo

Lee

et al. 2021

Front. Mol. Biosci.

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Behcet’s disease (BD) is an immune disease characterized by chronic and relapsing systemic vasculitis of unknown etiology, which can lead to blindness and even death. Despite continuous efforts to discover biomarkers for accurate and rapid diagnosis and optimal treatment of BD, there is still no signature marker with high sensitivity and high specificity. As the link between glycosylation and the immune system has been revealed, research on the immunological function of glycans is being actively conducted. In particular, sialic acids at the terminus of glycoconjugates are directly implicated in immune responses, cell–cell/pathogen interactions, and tumor progression. Therefore, changes in sialic acid epitope in the human body are spotlighted as a new indicator to monitor the onset and progression of immune diseases. Here, we performed global profiling of N-glycan compositions derived from the sera of 47 healthy donors and 47 BD patients using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) to preferentially determine BD target glycans. Then, three sialylated biantennary N-glycans were further subjected to the separation of linkage isomers and quantification using porous graphitized carbon-liquid chromatography (PGC-LC)/multiple reaction monitoring (MRM)-MS. We were able to successfully identify 11 isomers with sialic acid epitopes from the three glycan compositions consisting of Hex5HexNAc4NeuAc1, Hex5HexNAc4Fuc1NeuAc1, and Hex5HexNAc4NeuAc2. Among them, three isomers almost completely distinguished BD from control with high sensitivity and specificity with an area under the curve (AUC) of 0.945, suggesting the potential as novel BD biomarkers. In particular, it was confirmed that α2,3-sialic acid at the terminus of biantennary N-glycan was the epitope associated with BD. In this study, we present a novel approach to elucidating the association between BD and glycosylation by tracing isomeric structures containing sialic acid epitopes. Isomer-specific glycan profiling is suitable for analysis of large clinical cohorts and may facilitate the introduction of diagnostic assays for other immune diseases.

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Specific Analysis of α-2,3-Sialylated N-Glycan Linkage Isomers by Microchip Capillary Electrophoresis–Mass Spectrometry

Cited by 27 publications

References 48 publications

Fast Discrimination of Sialylated N-Glycan Linkage Isomers with One-Step Derivatization by Microfluidic Capillary Electrophoresis–Mass Spectrometry

Fast Discrimination of Sialylated N-Glycan Linkage Isomers with One-Step Derivatization by Microfluidic Capillary Electrophoresis–Mass Spectrometry

Advances in mass spectrometry‐based glycomics—An update covering the period 2017–2021

Isomer-Specific Monitoring of Sialylated N-Glycans Reveals Association of α2,3-Linked Sialic Acid Epitope With Behcet’s Disease

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