Separation of Permethylated O-Glycans, Free Oligosaccharides, and Glycosphingolipid-Glycans Using Porous Graphitized Carbon (PGC) Column

Cho, Byeong Gwan; Peng, Wenjing; Mechref, Yehia

doi:10.3390/metabo10110433

Cited by 21 publications

(16 citation statements)

References 45 publications

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“…An early study showed only partial separation of permethylated Man7 and Man8 isomers derived from RNase B ( Stavenhagen et al, 2015 ). Then, Zhou et al, 2017a , b introduced a high temperature PGC-LC-MS method that enables an efficient isomeric separation of both permethylated N-glycans and smaller glycans such as O-glycans, ganglioside glycans, and free oligosaccharide ( Cho et al, 2020 ). With the development of these methods, PGC-LC-MS of isomeric permethylated glycomics has been applied to address many biological issues such as breast cancer ( Peng et al, 2019 ), liver cancer ( Huang et al, 2017b ), colorectal cancer ( Madunić et al, 2021 ), and more ( Huang et al, 2017b ; Xu et al, 2020 ; Zhang et al, 2020b ; Flynn et al, 2021 ).…”

Section: Mass Spectrometry-based Separation Techniques Facilitate Gly...mentioning

confidence: 99%

MS-based glycomics: An analytical tool to assess nervous system diseases

et al. 2022

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Neurological diseases affect millions of peopleochemistryorldwide and are continuously increasing due to the globe’s aging population. Such diseases affect the nervous system and are characterized by a progressive decline in brain function and progressive cognitive impairment, decreasing the quality of life for those with the disease as well as for their families and loved ones. The increased burden of nervous system diseases demands a deeper insight into the biomolecular mechanisms at work during disease development in order to improve clinical diagnosis and drug design. Recently, evidence has related glycosylation to nervous system diseases. Glycosylation is a vital post-translational modification that mediates many biological functions, and aberrant glycosylation has been associated with a variety of diseases. Thus, the investigation of glycosylation in neurological diseases could provide novel biomarkers and information for disease pathology. During the last decades, many techniques have been developed for facilitation of reliable and efficient glycomic analysis. Among these, mass spectrometry (MS) is considered the most powerful tool for glycan analysis due to its high resolution, high sensitivity, and the ability to acquire adequate structural information for glycan identification. Along with MS, a variety of approaches and strategies are employed to enhance the MS-based identification and quantitation of glycans in neurological samples. Here, we review the advanced glycomic tools used in nervous system disease studies, including separation techniques prior to MS, fragmentation techniques in MS, and corresponding strategies. The glycan markers in common clinical nervous system diseases discovered by utilizing such MS-based glycomic tools are also summarized and discussed.

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Section: Mass Spectrometry-based Separation Techniques Facilitate Gly...mentioning

confidence: 99%

MS-based glycomics: An analytical tool to assess nervous system diseases

et al. 2022

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“…Exoglycosidase enzymatic digestion has been utilized to elucidate the glycan and glycopeptide isomers (Cho et al, 2020; Royle et al, 2006). However, this approach is commonly combined with LC‐MS or CE‐MS to separate the digested products from the precursor glycans.…”

Section: Ms‐based Isomeric Characterization Of Glycans and Glycopeptidesmentioning

confidence: 99%

“…Dong et al examined the reduced and permethylated free oligosaccharide isomers derived from human, bovine, and goat milk samples (Dong et al, 2016). Subsequently, PGC‐LC‐MS was utilized to demonstrate the isomeric separation of free reducing end and reduced end O‐ glycans, free oligosaccharides from human milk, and GSL glycans derived from the MDA‐MB‐231BR cancer cell line (Cho et al, 2020).…”

Section: Lc‐ms Analysis Of Permethylated Glycan Isomersmentioning

confidence: 99%

MS‐based glycomics and glycoproteomics methods enabling isomeric characterization

Peng

Reyes

Gautam

et al. 2021

Mass Spectrometry Reviews

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Glycosylation is one of the most significant and abundant posttranslational modifications in mammalian cells. It mediates a wide range of biofunctions, including cell adhesion, cell communication, immune cell trafficking, and protein stability. Also, aberrant glycosylation has been associated with various diseases such as diabetes, Alzheimer's disease, inflammation, immune deficiencies, congenital disorders, and cancers. The alterations in the distributions of glycan and glycopeptide isomers are involved in the development and progression of several human diseases. However, the microheterogeneity of glycosylation brings a great challenge to glycomic and glycoproteomic analysis, including the characterization of isomers. Over several decades, different methods and approaches have been developed to facilitate the characterization of glycan and glycopeptide isomers.Mass spectrometry (MS) has been a powerful tool utilized for glycomic and glycoproteomic isomeric analysis due to its high sensitivity and rich structural information using different fragmentation techniques. However, a comprehensive characterization of glycan and glycopeptide isomers remains a challenge when utilizing MS alone. Therefore, various separation methods, including liquid chromatography, capillary electrophoresis, and ion mobility, were developed to resolve glycan and glycopeptide isomers before MS. These separation techniques were coupled to MS for a better identification and quantitation of glycan and glycopeptide isomers. Additionally, bioinformatic tools are essential for the automated processing of glycan and glycopeptide isomeric data to facilitate isomeric studies in biological cohorts. Here in this review, we discuss commonly employed MS-based techniques, separation hyphenated MS methods, and software, facilitating the separation, identification, and quantitation of glycan and glycopeptide isomers.

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“…Compared to RPLC and HILIC, PGC has higher isomeric separation efficiency due to its retention mechanism. PGC columns have been widely employed as a powerful tool for glycan profiling and isomeric separation studies in native, reduced end, and permethylated glycans [84,93,105,106]. The hydrophobic interaction between the stationary phase and analytes, and the remarkable polar retention effect of graphite, facilitate the isomeric separation.…”

Section: Pgc Separationmentioning

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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Separation of Permethylated O-Glycans, Free Oligosaccharides, and Glycosphingolipid-Glycans Using Porous Graphitized Carbon (PGC) Column

Cited by 21 publications

References 45 publications

MS-based glycomics: An analytical tool to assess nervous system diseases

MS-based glycomics: An analytical tool to assess nervous system diseases

MS‐based glycomics and glycoproteomics methods enabling isomeric characterization

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

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