Recent advances in coupling capillary electrophoresis‐based separation techniques to ESI and MALDI‐MS

Zhong, Xuefei; Zhang, Zichuan; Jiang, Shan; Li, Lingjun

doi:10.1002/elps.201300451

Cited by 62 publications

(40 citation statements)

References 98 publications

(153 reference statements)

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“…Enzymatic studies 37 are sometimes inconclusive because the enzymes lack sufficient specificity and reactions do not always go to completion, which results in incomplete characterization of a set of structures. The direct interfacing of capillary electrophoresis with mass spectrometry (CE-MS) 41–45 brings together high resolution separations and unambiguous structural identification. Several commercial options now exist to couple CE instrumentation to mass spectrometers in a facile and reproducible manner.…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis–mass spectrometry for direct structural identification of serum N-glycans

Snyder

Zhou

Karty

et al. 2017

Journal of Chromatography A

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Through direct coupling of capillary electrophoresis (CE) to mass spectrometry (MS) with a sheathless interface, we have identified 77 potential N-glycan structures derived from human serum. We confirmed the presence of N-glycans previously identified by indirect methods, e.g., electrophoretic mobility standards, obtained 31 new N-glycan structures not identified in our prior work, differentiated co-migrating structures, and determined specific linkages on isomers featuring sialic acids. Serum N-glycans were cleaved from proteins, neutralized via methylamidation, and labeled with the fluorescent tag 8-aminopyrene-1,3,6-trisulfonic acid, which renders the glycan fluorescent and provides a -3 charge for electrophoresis and negative-mode MS detection. The neutralization reaction also stabilizes the labile sialic acids. In addition to methylamidation, native charges from sialic acids were neutralized through reaction with 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium to amidate α2,6-linked sialic acids in the presence of ammonium chloride and form lactones with α2,3-linked sialic acids. This neutralization effectively labels each type of sialic acid with a unique mass to determine specific linkages on sialylated N-glycans. For both neutralization schemes, we compared the results from microchip electrophoresis and CE.

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Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis–mass spectrometry for direct structural identification of serum N-glycans

Snyder

Zhou

Karty

et al. 2017

Journal of Chromatography A

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“…It features rapid analyte separations based on the electrophoretic mobility of molecules, including those with the same molecular weights (e.g., diastereomers), with high resolving power and low sample consumption (a microliter or less). 31,32 Many aspects of CE have greatly progressed in recent decades, and include the development of advanced separation modes and nanoscale sampling, and the interface of CE with different detection methods. 30,67 While CE is powerful on its own, it is even more productive when coupled with optical, electrochemical, or MS-based detection.…”

Section: Specific Cell-type Targeting: Meeting the Needs For Separatimentioning

confidence: 99%

“…Interdisciplinary developments include single-cell genomics and transcriptomics, 16−19 electrochemistry, 20−22 single-molecule microscopy and spectroscopy, 23−26 nuclear magnetic resonance, 27,28 capillary electrophoresis (CE), 29−32 MS, 6,33−37 and microfluidics, 38,39 to name a few. Clearly, single-cell “omics” comprises a number of rapidly growing interdisciplinary fields.…”

Section: Introductionmentioning

confidence: 99%

Categorizing Cells on the Basis of their Chemical Profiles: Progress in Single-Cell Mass Spectrometry

et al. 2017

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The chemical differences between individual cells within large cellular populations provide unique information on organisms’ homeostasis and the development of diseased states. Even genetically identical cell lineages diverge due to local microenvironments and stochastic processes. The minute sample volumes and low abundance of some constituents in cells hinder our understanding of cellular heterogeneity. Although amplification methods facilitate single-cell genomics and transcriptomics, the characterization of metabolites and proteins remains challenging both because of the lack of effective amplification approaches and the wide diversity in cellular constituents. Mass spectrometry has become an enabling technology for the investigation of individual cellular metabolite profiles with its exquisite sensitivity, large dynamic range, and ability to characterize hundreds to thousands of compounds. While advances in instrumentation have improved figures of merit, acquiring measurements at high throughput and sampling from large populations of cells are still not routine. In this Perspective, we highlight the current trends and progress in mass-spectrometry-based analysis of single cells, with a focus on the technologies that will enable the next generation of single-cell measurements.

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“…LIF detection is often used when low internal diameters are employed because of the inherent sensitivity of fluorescence detection. Despite inherent challenges in coupling electrophoresis to MS recent advances in MS sensitivity[18] may eventually allow detection by this label-free, highly selective detector.…”

Section: Improvements To Throughputmentioning

confidence: 99%

Advances in capillary electrophoresis and the implications for drug discovery

Ouimet

D'Amico

Kennedy

2016

Expert Opinion on Drug Discovery

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Introduction Many screening platforms are prone to assay interferences that can be avoided by directly measuring the target or enzymatic product. Capillary electrophoresis (CE) and microchip electrophoresis (MCE) have been applied in a variety of formats to drug discovery. CE provides direct detection of the product allowing for the identification of some forms of assay interference. The high efficiency, rapid separations, and low volume requirements make CE amenable to drug discovery. Areas Covered This article describes advances in capillary electrophoresis throughput, sample introduction, and target assays as they pertain to drug discovery and screening. Instrumental advances discussed include integrated droplet microfluidics platforms and multiplexed arrays. Applications of CE to assays of diverse drug discovery targets, including enzymes and affinity interactions are also described. Expert opinion Current screening with CE does not fully take advantage of the throughputs or low sample volumes possible with CE and is most suitable as a secondary screening method or for screens that are inaccessible with more common platforms. With further development, droplet microfluidics coupled to MCE could take advantage of the low sample requirements by performing assays on the nanoliter scale at high throughput.

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Recent advances in coupling capillary electrophoresis‐based separation techniques to ESI and MALDI‐MS

Cited by 62 publications

References 98 publications

Capillary electrophoresis–mass spectrometry for direct structural identification of serum N-glycans

Capillary electrophoresis–mass spectrometry for direct structural identification of serum N-glycans

Categorizing Cells on the Basis of their Chemical Profiles: Progress in Single-Cell Mass Spectrometry

Advances in capillary electrophoresis and the implications for drug discovery

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