Using micro pillar array columns (μPAC) for the analysis of permethylated glycans

Cho, Byeong Gwan; Jiang, Peilin; Goli, Mona; Gautam, Sakshi; Mechref, Yehia

doi:10.1039/d1an00643f

Cited by 16 publications

(16 citation statements)

References 42 publications

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“…In the last few years, micro-array pillar nano-LC (µPAC) columns have been commercialized that have favorable properties like good reproducibility, durability, and low back pressure. Recently, Cho et al [ 121 ] separated the permethylated N- and O-glycans of RNase B and fetuin on a 50 or 200 cm µPAC column (5.0 μm pillar diameter, 2.5 µm inter pillar distance, 18 µm pillar height, 50 or 200 cm bed length). In this work, they used a standard nano-LC Acclaim PepMap ™ C18 column (0.075 × 150 mm; 2.0 µm), too.…”

Section: Reversed Phase Chromatographymentioning

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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Section: Reversed Phase Chromatographymentioning

confidence: 99%

Liquid chromatography and capillary electrophoresis in glycomic and glycoproteomic analysis

et al. 2022

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“…With the pillar array system, the coefficient of variation for the peptide retention times was improved 2.5-fold, and high interlaboratory reproducibility was achieved compared to conventional LC. Moreover, pillar array columns (50 cm) were effective for separating some permethylated glycan isomers [36].…”

Section: Pillar Arraymentioning

confidence: 99%

On-Chip Liquid Chromatography

Tsunoda

2022

Encyclopedia

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On-chip liquid chromatography (LC) refers to LC technology that is miniaturized to fit on a microchip to enable fast, high-throughput analysis, with small sample volumes and low reagent consumption. Four different on-chip LC approaches have been developed to date: use of open-tubular, packed-particle, monolithic, and pillar array columns. These methods have been applied to proteomics as well as the analysis of small molecules and drugs in various biological samples. Recent advances in on-chip LC are summarized herein.

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“…While bed‐packed RP columns are commonly employed for permethylated glycan separations, the use of newly commercialized micro array pillar nanoLC columns (μPAC, Pharmafluidics) has not been demonstrated for glycan analysis [94]. The uniformly employed micropillars, coated with C18 material in μPAC, present an efficient and robust RP‐LC platform.…”

Section: Recent Advances In Separation Techniquesmentioning

confidence: 99%

“…This column has been established in proteomics analysis [95, 96]; however, Cho et al. [94] recently developed the workflow of μPAC to analyze reduced and permethylated glycans. The work assessed the ability of 50 cm μPAC to analyze both permethylated N ‐ and O ‐glycans.…”

Section: Recent Advances In Separation Techniquesmentioning

confidence: 99%

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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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Using micro pillar array columns (μPAC) for the analysis of permethylated glycans

Abstract: Glycosylation is a complex and common post-translational modification of proteins. To study glycosylation, liquid chromatography-mass spectrometry (LC-MS) is often used to profile and structurally characterize the glycans in biological systems....

Cited by 16 publications

References 42 publications

Liquid chromatography and capillary electrophoresis in glycomic and glycoproteomic analysis

Liquid chromatography and capillary electrophoresis in glycomic and glycoproteomic analysis

On-Chip Liquid Chromatography

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

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