Ultrafast high-resolution analysis of human milk oligosaccharides by multicapillary gel electrophoresis

Sarkozy, Daniel; Borza, Beáta; Domokos, Apolka; Váradi, Eszter; Szigeti, Márton; Meszaros-Matwiejuk, Agnes; Molnár-Gábor, Dóra; Guttman, András

doi:10.1016/j.foodchem.2020.128200

Cited by 21 publications

(15 citation statements)

References 30 publications

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“…Finally, C(G)E-based methods were also successfully applied for the absolute quantification of carbohydrates (Sarkozy et al 2021;Eussen et al 2021), which completes the wide field of applications. Taken together, unprecedented separation together with low costs, robustness, speed of analysis, multiplexing capability, high sensitivity, and nano-to femtoliter injection volumes make CE a veritable competitor to other more traditionally used techniques for glycan analysis.…”

Section: Background: Principles Of Capillary (Gel) Electrophoresis (C(g)e)mentioning

confidence: 92%

Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Cajic

Hennig

Burock³

et al. 2021

Experientia Supplementum

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The in-depth characterization of protein glycosylation has become indispensable in many research fields and in the biopharmaceutical industry. Especially knowledge about modulations in immunoglobulin G (IgG) N-glycosylation and their effect on immunity enabled a better understanding of human diseases and the development of new, more effective drugs for their treatment. This chapter provides a deeper insight into capillary (gel) electrophoresis-based (C(G)E) glycan analysis, addressing its impressive performance and possibilities, its great potential regarding real high-throughput for large cohort studies, as well as its challenges and limitations. We focus on the latest developments with respect to miniaturization and mass spectrometry coupling, as well as data analysis and interpretation. The use of exoglycosidase sequencing in combination with current C(G)E technology is discussed, highlighting possible difficulties and pitfalls. The application section describes the detailed characterization of N-glycosylation, utilizing multiplexed CGE with laser-induced fluorescence detection (xCGE-LIF). Besides a comprehensive overview on antibody glycosylation by comparing species-specific IgGs and human immunoglobulins A, D, E, G, and M, the chapter comprises a comparison of therapeutic monoclonal antibodies from different production cell lines, as well as a detailed characterization of Fab and Fc glycosylation. These examples illustrate the full potential of C(G)E, resolving the smallest differences in sugar composition and structure.

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Section: Background: Principles Of Capillary (Gel) Electrophoresis (C(g)e)mentioning

confidence: 92%

Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Cajic

Hennig

Burock³

et al. 2021

Experientia Supplementum

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“…Although the involved enzymes are not described, it is likely by tight sequential addition of GlcNAc and Gal units. The intermediate lacto‐ N ‐triose with only a GlcNAc added to lactose is only rarely reported in human milk 15,16 . Both LNT and LNnT as well as their further LNB‐ and LacNAc‐elongated descendants can be decorated with Fuc and NeuAc.…”

Section: What Are Hmos?mentioning

confidence: 99%

“…The intermediate lacto‐ N ‐triose with only a GlcNAc added to lactose is only rarely reported in human milk. 15 , 16 Both LNT and LNnT as well as their further LNB‐ and LacNAc‐elongated descendants can be decorated with Fuc and NeuAc. Because many GTs are involved in the formation of several linkage types, a large variety of structures differing in composition, conformation and chain length is found in human milk.…”

Section: What Are Hmos?mentioning

confidence: 99%

Biology of human milk oligosaccharides: From basic science to clinical evidence

Sprenger

Tytgat

Binia

et al. 2022

J Human Nutrition Diet

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Human milk oligosaccharides (HMOs) have been researched by scientists for over 100 years, driven by the substantial evidence for the nutritional and health benefits of mother's milk. Yet research has truly bloomed during the last decade, thanks to progress in biotechnology, which has allowed the production of large amounts of bona fide HMOs. The availability of HMOs has been particularly crucial for the renewed interest in HMO research because of the low abundance or even absence of HMOs in farmed animal milk. This interest is reflected in the increasing number of original research publications and reviews on HMOs. Here, we provide an overview and critical discussion on structure–function relations of HMOs that highlight why they are such interesting and important components of human milk. Clinical observations in breastfed infants backed by basic research from animal models provide guidance as to what physiological roles for HMOs are to be expected. From an evidence‐based nutrition viewpoint, we discuss the current data supporting the clinical relevance of specific HMOs based on randomised placebo‐controlled clinical intervention trials in formula‐fed infants.

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“…Different gel compositions have been studied to this aim. For instance, the potential performance of borate-buffered dextran gel and C100HT separation gel buffer-buffer system (conventional type carbohydrate separation matrix) was compared for the separation of 10 major HMOs after derivatization with aminopyrenetrisulfonate fluorescent label [73]. Twelve capillaries were employed simultaneously, achieving LODs and LOQs as low as 0.025 mg/mL and 0.100 mg/mL, respectively.…”

Section: Carbohydratesmentioning

confidence: 99%

Capillary electromigration methods for food analysis and Foodomics: Advances and applications in the period February 2019–February 2021

et al. 2021

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This work presents a revision of the main applications of capillary electromigration methods in food analysis and Foodomics. Articles that were published during the period February 2019‐February 2021 are included. The work shows the multiple CE methods that have been developed and applied to analyze different types of molecules in foods. Namely, CE methods have been applied to analyze amino acids, biogenic amines, carbohydrates, chiral compounds, contaminants, DNAs, food additives, heterocyclic amines, lipids, secondary metabolites, peptides, pesticides, phenols, pigments, polyphenols, proteins, residues, toxins, vitamins, small organic and inorganic compounds, as well as other minor compounds. The last results on the use of CE for monitoring food interactions and food processing, including recent microchips developments and new applications of CE in Foodomics, are discussed too. The new procedures of CE to investigate food quality and safety, nutritional value, storage and bioactivity are also included in the present review work.

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Ultrafast high-resolution analysis of human milk oligosaccharides by multicapillary gel electrophoresis

Cited by 21 publications

References 30 publications

Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Biology of human milk oligosaccharides: From basic science to clinical evidence

Capillary electromigration methods for food analysis and Foodomics: Advances and applications in the period February 2019–February 2021

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