Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography–electrospray ionization-mass spectrometry with graphitic carbon stationary phase

Michael, Claudia; Rizzi, Andreas

doi:10.1016/j.chroma.2015.01.028

Cited by 28 publications

(16 citation statements)

References 43 publications

(51 reference statements)

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“…LC‐MS/MS is currently considered a great tool for biomolecule identification, enabling simultaneous glycan isomeric separation and structural assignment. Various separation techniques have been developed to resolve glycan isomers at both the glycan and glycopeptide levels . Hydrophilic liquid interaction chromatography (HILIC) coupled with reducing end labeling or derivatization is an excellent approach to separate glycans or glycopeptides .…”

Section: Introductionmentioning

confidence: 99%

LC‐MS/MS isomeric profiling of permethylated N‐glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients

et al. 2017

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Early stage detection and cancer treatment demand the identification of reliable biomarkers. Over the past decades, efforts have been devoted to assess the variation of glycosylation level as well as the glycan structures of proteins in blood or serum, associated with the development and/or progression of several cancers, including liver. Herein, an LC-MS/MS-based analysis was conducted to define the glycosylation patterns of haptoglobin glycoprotein derived from sera collected from cirrhotic and hepatocellular carcinoma (HCC) patients. The haptoglobin samples were extracted from serum using an antibody-immobilized column prior to the release of N-glycans. A comparison of non-isomeric and isomeric permethylated glycan forms was achieved using C18 and porous graphitic carbon (PGC) columns, respectively. In the case of C18-LC-MS/MS analysis, 25 glycan structures were identified of which 10 sialylated structures were found to be statistically significant between the two cohorts. Also, 8 out of 34 glycan structures identified by PGC-LC-MS/MS were found to be statistically significant, suggesting that isomeric distributions of a particular glycan composition were different in abundances between the two cohorts. The glycan isoform patterns distinguished early stage HCC from cirrhotic patients. Both retention times and tandem mass spectra were utilized to determine the specific isomeric glycan structures. All of the glycan isomers, which were statistically significant, were either branch fucosylated or composed of α-2,6 linked sialic acid moieties. The result of this study demonstrates the potential importance of isomeric separation for defining disease prompted aberrant glycan changes. The levels of several glycan isomers effectively distinguished early stage HCC from cirrhosis.

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Section: Introductionmentioning

confidence: 99%

LC‐MS/MS isomeric profiling of permethylated N‐glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients

et al. 2017

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“…A more comprehensive discussion on PGC supports is available 30 , as is a more exhaustive discussion on comparisons between PGC supports versus reversed-phase and HILIC ones for glycan analysis 31 . Since PGC columns remain attractive for polar analytes, these packings have seen use in the separation of glycans isolated from mammalian sources 32,33 and have applications in the separation of other carbohydrates such as underivatized oligosaccharide anomers in vegetable matrices 34 , cello-oligosaccharides 35,36 and fructan mixtures 37 . While these PGC packings remain somewhat unexplored for carbohydrate purification, a few drawbacks associated with these supports have come to light, including the loss of retention/variability in retention times 31,38 and poor stability of the packing material under acidic mobile phase conditions 39 .…”

Section: Analytical Toolbox Of Chromatographic Modesmentioning

confidence: 99%

Recent liquid chromatographic approaches and developments for the separation and purification of carbohydrates

2017

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“…106–108 Additional strategies include the use of the reducing-end label Girard’s Reagent P in deuterated or nondeuterated form 114 or reductive amination using 12 C 6 -aniline and 13 C 6 -aniline as isotope-coded labeling reagents. 115,116 …”

Section: Quantification Of Glycansmentioning

confidence: 99%

Recent Advances in the Analysis of Complex Glycoproteins

et al. 2016

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Quantitative isomer-specific N-glycan fingerprinting using isotope coded labeling and high performance liquid chromatography–electrospray ionization-mass spectrometry with graphitic carbon stationary phase

Cited by 28 publications

References 43 publications

LC‐MS/MS isomeric profiling of permethylated N‐glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients

LC‐MS/MS isomeric profiling of permethylated N‐glycans derived from serum haptoglobin of hepatocellular carcinoma (HCC) and cirrhotic patients

Recent liquid chromatographic approaches and developments for the separation and purification of carbohydrates

Recent Advances in the Analysis of Complex Glycoproteins

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