High-throughput and high-sensitivity N-Glycan profiling: A platform for biopharmaceutical development and disease biomarker discovery

Xie, Yongjing; Mota, Letícia Martins; Bergin, Adam; O’Flaherty, Róisín; Jones, Aled Wynne; Morgan, Bethan; Butler, Michael

doi:10.1016/j.ab.2021.114205

Cited by 14 publications

(19 citation statements)

References 76 publications

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“…In the current study, we extended our previously described high-throughput, high-sensitivity N-glycan preparation platform ( Xie et al. 2021 ) further to identify potential alterations in human serum N-glycome upon or after infection with SARS-CoV-2.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, the development of high-throughput and high-sensitivity technologies for reliable analysis of human serum glycomic profile can be a valuable tool in the study of disease or viral infection. In the current study, we extended our previously developed 96-well-plate-based high-throughput, high-sensitivity glycan preparation platform ( Xie et al. 2021 ), and related serum N-glycan identification and analytical method ( Xie and Butler 2022 ) even further to COVID-19 research with the aim of elucidating any potential COVID-19-associated serum N-glycomic changes and consequently gain better understand of this disease.…”

Section: Introductionmentioning

confidence: 99%

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Serum N-glycomic profiling may provide potential signatures for surveillance of COVID-19

Xie

Butler

2022

Glycobiology

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Disease development and progression are often associated with aberrant glycosylation, indicating that changes in biological fluid glycome may potentially serve as disease signatures. The corona virus disease-2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) represents a significant threat to global human health. However, the effect of SARS-CoV-2 infection on the overall serum N-glycomic profile has been largely unexplored. Here, we extended our 96-well-plate-based high-throughput, high-sensitivity N-glycan profiling platform further with the aim of elucidating potential COVID-19-associated serum N-glycomic alterations. Use of this platform revealed both similarities and differences between the serum N-glycomic fingerprints of COVID-19 positive and control cohorts. Although there were no specific glycan peaks exclusively present or absent in COVID-19 positive cohort, this cohort showed significantly higher levels of glycans and variability. On the contrary, the overall N-glycomic profiles for healthy controls were well-contained within a narrow range. From the serum glycomic analysis, we were able to deduce changes in different glycan subclasses sharing certain structural features. Of significance was the hyperbranched and hypersialylated glycans and their derived glycan subclass traits. T-distributed stochastic neighbour embedding (tSNE) and hierarchical heatmap clustering analysis were performed to identify 13 serum glycomic variables that potentially distinguished the COVID-19 positive from healthy controls. Such serum N-glycomic changes described herein may indicate or correlate to the changes in serum glycoproteins upon COVID-19 infection. Furthermore, mapping the serum N-glycome following SARS-CoV-2 infection may help us better understand the disease and enable “Long-COVID” surveillance to capture the full spectrum of persistent symptoms.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Serum N-glycomic profiling may provide potential signatures for surveillance of COVID-19

Xie

Butler

2022

Glycobiology

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“…Xie et al. [71] examined the performance of the InstantPC, ProA, and 2‐AB on N ‐glycans of rituximab using HILIC‐FLD. They found that InstantPC provided improvements of 1.3‐ and 100‐fold in FLR intensity than ProA and 2‐AB, respectively.…”

Section: Derivatization Of Glycansmentioning

confidence: 99%

“…Advantages of these tools include improved ionization from tertiary amine groups and enhanced fluorescent intensity from fluorophores. Recently, a high‐throughput platform for a fast glycan profile was generated using an InstantPC kit [71]. This rapid approach includes a rapid glycoprotein denaturation, deglycosylation, and fluorescent derivation by activating carbamate chemistry.…”

Section: Derivatization Of Glycansmentioning

confidence: 99%

“…Labeling efficiencies of the three labels were comparable with each other while relatively poorer repeatability was observed in RFMS-labeled glycans, especially for some disialylated ones. Xie et al [71] examined the performance of the InstantPC, ProA, and 2-AB on N-glycans of rituximab using HILIC-FLD. They found that InstantPC provided improvements of 1.3-and 100-fold in FLR intensity than ProA and 2-AB, respectively.…”

Section: Comparing Different Derivatization 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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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

Harvey

2024

Mass Spectrometry Reviews

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The use of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry for the analysis of carbohydrates and glycoconjugates is a well‐established technique and this review is the 12th update of the original article published in 1999 and brings coverage of the literature to the end of 2022. As with previous review, this review also includes a few papers that describe methods appropriate to analysis by MALDI, such as sample preparation, even though the ionization method is not MALDI. The review follows the same format as previous reviews. It is divided into three sections: (1) general aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, fragmentation, quantification and the use of computer software for structural identification. (2) Applications to various structural types such as oligo‐ and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals, and (3) other general areas such as medicine, industrial processes, natural products and glycan synthesis where MALDI is extensively used. Much of the material relating to applications is presented in tabular form. MALDI is still an ideal technique for carbohydrate analysis, particularly in its ability to produce single ions from each analyte and advancements in the technique and range of applications show little sign of diminishing.

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High-throughput and high-sensitivity N-Glycan profiling: A platform for biopharmaceutical development and disease biomarker discovery

Cited by 14 publications

References 76 publications

Serum N-glycomic profiling may provide potential signatures for surveillance of COVID-19

Serum N-glycomic profiling may provide potential signatures for surveillance of COVID-19

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

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

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