Quantitative GSL-glycome analysis of human whole serum based on an EGCase digestion and glycoblotting method

Furukawa, Yoshihiro; Sakaï, Shota; Yokota, Ikuko; Okada, Kazue; Hanamatsu, Hisatoshi; Kobayashi, Takashi; Yoshida, Yukuo; Higashino, Kenichi; Tamura, Tomohiro; Igarashi, Yasuyuki; Shinohara, Yasuro

doi:10.1194/jlr.d062083

Cited by 27 publications

(65 citation statements)

References 28 publications

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“…As some low‐molecular‐weight glycans (e.g. Hex 2 of GSL‐glycans, Hex 1 HexNAc 1 of fOSs) were partially lost during the solid‐phase extraction process, the absolute quantity was estimated by applying correction factors (Supporting Information Table 1) . Regarding bis‐PMP‐labeled glycans, we also observed fairly constant peak areas regardless of the differences in structure and molecular weight .…”

Section: Methodsmentioning

confidence: 80%

“…In glycoblotting, glycans were subjected to on‐bead methyl esterification to obtain sialylated oligosaccharides that were the chemical equivalents of neutral oligosaccharides . As we previously reported , signal strengths of a mixture of equal quantities of 14 aoWR labeled glycans with molecular weights ranging from 772 to 2826 were similar, indicating that the structure and molecular weight have no significant effect on the signal strength. As some low‐molecular‐weight glycans (e.g.…”

Section: Methodsmentioning

confidence: 89%

“…We recently extensively analyzed the GSL‐glycomes in human serum . In this study, we compared the serum GSL‐glycomes between human and mouse (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…A total of 20 μL of serum was delipidated and digested to GAG disaccharides. Detailed procedures and materials are provided elsewhere .…”

Section: Methodsmentioning

confidence: 99%

“…Purified N‐glycans, fOSs, O‐glycans, and GSL‐glycan solutions were mixed with 2,5‐dihydrobenzoic acid solution and subjected to MALDI‐TOF‐MS analysis as previously described . All peaks were selected using FlexAnalysis 3.0 with the SNAP algorithm, which fits isotopic patterns to the matching experimental data.…”

Section: Methodsmentioning

confidence: 99%

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Quantitative analysis of total serum glycome in human and mouse

et al. 2016

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Model mice are frequently used in drug discovery research. Knowledge of similarities and differences between the mouse and human glycomes is critical when model mice are used for the discovery of glycan-related biomarkers and targets for therapeutic intervention. Since few comparative glycomic studies between human and mouse have been conducted, we performed a comprehensive comparison of the major classes of glycans in human and mouse sera using mass spectrometric and liquid chromatographic analyses. Up to 131 serum glycans, including N-glycans, free oligosaccharides (fOSs), glycosaminoglycans, O-glycans, and glycosphingolipid (GSL)-glycans, were quantified. In both serum samples, N-glycans were the most abundant in the total serum glycome, while fOSs were the least abundant. As expected, the diversity of sialic acid (i.e. Neu5Ac vs. Neu5Gc) was the major species difference between human and mouse in terms of N- and O-glycosylation, while GSL-glycomic profiles were completely different, even when the sialic acid diversity was taken into consideration. Furthermore, total serum glycomics of STAM mouse were unveiled as an initial step to identify novel biomarkers of liver diseases, with which we could identify several glycans with expression significantly increased or decreased expression.

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Section: Methodsmentioning

confidence: 80%

Section: Methodsmentioning

confidence: 89%

“…We recently extensively analyzed the GSL‐glycomes in human serum . In this study, we compared the serum GSL‐glycomes between human and mouse (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…A total of 20 μL of serum was delipidated and digested to GAG disaccharides. Detailed procedures and materials are provided elsewhere .…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Quantitative analysis of total serum glycome in human and mouse

et al. 2016

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Analysis of Carbohydrates and Glycoconjugates by Matrix‐assisted Laser Desorption/Ionization Mass Spectrometry: An Update for 2015–2016

Harvey

2021

Mass Spectrometry Reviews

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This review is the ninth update of the original article published in 1999 on the application of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry to the analysis of carbohydrates and glycoconjugates and brings coverage of the literature to the end of 2016. Also included are papers that describe methods appropriate to analysis by MALDI, such as sample preparation techniques, even though the ionization method is not MALDI. Topics covered in the first part of the review include general aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, fragmentation and arrays. The second part of the review is devoted to applications to various structural types such as oligo‐ and poly‐saccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals. Much of this material is presented in tabular form. The third part of the review covers medical and industrial applications of the technique, studies of enzyme reactions and applications to chemical synthesis. The reported work shows increasing use of combined new techniques such as ion mobility and the enormous impact that MALDI imaging is having. MALDI, although invented over 30 years ago is still an ideal technique for carbohydrate analysis and advancements in the technique and range of applications show no sign of deminishing. © 2020 Wiley Periodicals, Inc.

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Comprehensive Cellular Glycan Profiling of Glycoproteins and Glycosphingolipids by Glycoblotting and BEP Methods

Hanamatsu

Furukawa

2022

Methods in Molecular Biology

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Quantitative GSL-glycome analysis of human whole serum based on an EGCase digestion and glycoblotting method

Cited by 27 publications

References 28 publications

Quantitative analysis of total serum glycome in human and mouse

Quantitative analysis of total serum glycome in human and mouse

Analysis of Carbohydrates and Glycoconjugates by Matrix‐assisted Laser Desorption/Ionization Mass Spectrometry: An Update for 2015–2016

Comprehensive Cellular Glycan Profiling of Glycoproteins and Glycosphingolipids by Glycoblotting and BEP Methods

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