High sensitivity capillary electrophoresis with fluorescent detection for glycan mapping

Liénard--Mayor, Théo; Yang, Bin; Tran, Nguyet Thuy; Bruneel, Arnaud; Guttman, András; Taverna, Myriam; Duc, Thanh

doi:10.1016/j.chroma.2021.462593

Cited by 15 publications

(7 citation statements)

References 42 publications

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“…These IgG glycans are known to be mainly biantennary complex glycans with low terminal sialylation (5-10%) and high core fucosylation (> 92%), with a small proportion containing a bisecting GlcNAc (11%)[43]. The CE-LIF profile of IgG glycans obtained with our approach is very similar to those already reported that used conventional tube-based protocols and CE profiling[36,44]. From the electropherograms obtained for human IgG, the relative proportion of the five most abundant glycans was calculated and compared to those obtained with in-tube labeling from the literature (Table2).…”

supporting

confidence: 76%

“…Analyses of APTS‐labeled glycans or malto‐oligosaccharides were carried out with our recently developed BGE composed of triethanolamine/citric acid (IS of 100 mM, pH 4.75) [36]. These separations were implemented using a fused‐silica capillary of 50 μm id, with a total length (L tot ) of 60 cm and an effective length (L eff ) of 50 cm, under a separation voltage of −25 kV.…”

Section: Methodsmentioning

confidence: 99%

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In capillary labeling and online electrophoretic separation of N‐glycans from glycoproteins

Yang

Duc

Tran

et al. 2022

J of Separation Science

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In this study, we present a new approach for in-capillary fluorescent labeling of N-glycans prior to their analysis with CE coupled with laser-induced fluorescent detection. This integrated approach allows using a CE capillary as a microreactor to perform several steps required for labeling glycans with 8aminopyrene-1,3,6 trisulfonic acid and at the same time as a separation channel for CE of fluorescently labeled glycans. This could be achieved through careful optimization of all different steps, including sequential injections of fluorescent dye and glycan plugs, mixing by transverse diffusion of laminar flow profiles, incubation in a thermostatic zone, and finally separation and detection with CE. Such a complex sample treatment protocol for glycan labeling that is feasible thus far only in batchwise mode can now be converted into an automated and integrated protocol. Our approach was applied successfully to analyze fluorescently labeled N-linked oligosaccharides released from human immunoglobulin G and rituximab, a monoclonal antibody used for cancer treatment. We demonstrated the superiority of this in-capillary approach over the conventional in-tube protocol, with fourfold less reagent consumption and full automation without remarkable degradation of the glycan separation profile obtained by capillary electrophoresis.

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supporting

confidence: 76%

Section: Methodsmentioning

confidence: 99%

In capillary labeling and online electrophoretic separation of N‐glycans from glycoproteins

Yang

Duc

Tran

et al. 2022

J of Separation Science

Self Cite

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“…Detection sensitivity is one of the main challenges in CE-LIF glycomic analysis. Liénard-Mayor et al [ 166 ] presented a novel strategy to improve detection sensitivity and peak capacity in CE-LIF analysis of APTS-labeled glycans. From the set of background electrolytes tested, they chose triethanolamine/citric acid and triethanolamine/acetic acid at high ionic strengths (up to 200 mM), which helped to create a large conductivity difference between sample zone and background electrolyte.…”

Section: Glycan Separationmentioning

confidence: 99%

Liquid chromatography and capillary electrophoresis in glycomic and glycoproteomic analysis

et al. 2022

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Glycosylation is one of the most significant and abundant post-translational modifications in cells. Glycomic and glycoproteomic analyses involve the characterization of oligosaccharides (glycans) conjugated to proteins. Glycomic and glycoproteomic analysis is highly challenging because of the large diversity of structures, low abundance, site-specific heterogeneity, and poor ionization efficiency of glycans and glycopeptides in mass spectrometry (MS). MS is a key tool for characterization of glycans and glycopeptides. However, MS alone does not always provide full structural and quantitative information for many reasons, and thus MS is combined with some separation technique. This review focuses on the role of separation techniques used in glycomic and glycoproteomic analyses, liquid chromatography and capillary electrophoresis. The most important separation conditions and results are presented and discussed. Graphical abstract

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“…A large-volume sample stacking with an EOF pump (LVSEP) is a sample preconcentration technique based on LVSS without PS, which allows both the sample enrichment and the removal of a vacant sample matrix under a constant voltage. Recently, LVSEP has been applied to the oligosaccharide [47], amine [48], and glycosaminoglycan [49] analyses. Kitagawa et al combined LVSEP with FASI for the sensitive determination of a standard fluorescent dye and amino acid enantiomers [50].…”

Section: Application Of Field-amplified Sample Stackingmentioning

confidence: 99%

Recent Applications of Dynamic On-Line Sample Preconcentration Techniques in Capillary Electrophoresis

Kitagawa

SOMA

2022

Chromatography

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This review highlights recent applications of on-line sample preconcentration techniques in capillary electrophoresis (CE) from 2019 to 2021. Various preconcentration techniques based on the analyte velocity change in two or three discontinuous solutions system including field-amplified sample stacking, transient isotachophoresis, pH-mediated stacking, sweeping, and their modified and combined techniques have been employed to enrich and separate biological, environmental, food samples, and so on, in CE. More than 60 published research articles collected from Scopus databases described the dynamic on-line sample preconcentration techniques. This review provides comprehensive tables listing the applications of the on-line sample preconcentration techniques.

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High sensitivity capillary electrophoresis with fluorescent detection for glycan mapping

Cited by 15 publications

References 42 publications

In capillary labeling and online electrophoretic separation of N‐glycans from glycoproteins

In capillary labeling and online electrophoretic separation of N‐glycans from glycoproteins

Liquid chromatography and capillary electrophoresis in glycomic and glycoproteomic analysis

Recent Applications of Dynamic On-Line Sample Preconcentration Techniques in Capillary Electrophoresis

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