Comparison of the Human and Bovine Milk N-Glycome via High-Performance Microfluidic Chip Liquid Chromatography and Tandem Mass Spectrometry (original) (raw)
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Journal of agricultural …, 2011
N-Linked glycans of skim human milk proteins were determined for three mothers. N-Linked glycans are linked to immune defense, cell growth, and cellÀcell adhesion, but their functions in human milk are undetermined. Protein-bound N-linked glycans were released with peptidyl N-glycosidase F (PNGase F), enriched by graphitized carbon chromatography, and analyzed with Chip-TOF MS. To be defined as N-glycans, compounds were required, in all three procedural replicates, to match, within 6 ppm, against a theoretical human N-glycan library and be at least 2-fold higher in abundance in PNGase F-treated than in control samples. Fifty-two N-linked glycan compositions were identified, and 24 were confirmed via tandem mass spectra analysis. Twentyseven compositions have been found previously in human milk, and 25 are novel compositions. By abundance, 84% of N-glycans were fucosylated and 47% were sialylated. The majority (70%) of total N-glycan abundance was composed of N-glycans found in all three milk samples.
Composition and structure elucidation of human milk glycosaminoglycans
Glycobiology, 2011
To date, there is no complete structural characterization of human milk glycosaminoglycans (GAGs) available nor do any data exist on their composition in bovine milk. Total GAGs were determined on extracts from human and bovine milk. Samples were subjected to digestion with specific enzymes, treated with nitrous acid, and analyzed by agarose-gel electrophoresis and high-performance liquid chromatography for their structural characterization. Quantitative analyses yielded 7 times more GAGs in human milk than in bovine milk. In particular, galactosaminoglycans, chondroitin sulfate (CS) and dermatan sulfate (DS), were found to differ considerably from one type of milk to the other. In fact, hardly any DS was observed in human milk, but a low-sulfated CS having a very low charge density of 0.36 was found. On the contrary, bovine milk galactosaminoglycans were demonstrated to be composed of 66% DS and 34% CS for a total charge density of 0.94. Structural analysis performed by heparinases showed a prevalence of fast-moving heparin over heparan sulfate, accounting for 30-40% of total GAGs in both milk samples and showing lower sulfation in human (2.03) compared with bovine (2.28). Hyaluronic acid was found in minor amounts. This study offers the first full characterization of the GAGs in human milk, providing useful data to gain a better understanding of their physiological role, as well as of their fundamental contribution to the health of the newborn.
Recent advances in human milk glycobiology
Pediatric Research, 2014
The field of human milk glycobiology is progressing rapidly, with important potential applications to health. Only a few decades ago, most experts considered human milk oligosaccharides (HMOS) to be an incidental consequence of high concentrations of Users may view, print, copy, download and text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com
Human milk glycosaminoglycan composition from women of different countries: a pilot study
The Journal of Maternal-Fetal & Neonatal Medicine, 2019
Objective: In this pilot study we report the composition, structure and properties of glycosaminoglycans present in milk samples of various countries and ethnicities. Methods:50 samples of human milk were analysed, 10 from East Europe, 10 from North Africa, 10 from Central Africa, 10 from South America and 10 from Asia. Moreover, 30 samples were obtained during the first week and 20 between 8 to 30 days of life. Results: Overall, no significant differences were observed for the qualitative composition of GAGs, mainly chondroitin sulfate, heparan sulfate and hyaluronic acid, comparing the mothers from the various countries and between the 30 milks obtained during the first week and the 20 samples collected thereafter. Moreover, no significant differences in human milk GAGs within the different groups analysed belonging to various counties and ethnicities were observed. Conclusions:These results may be of useful, as in the case of pilot studies with infant formulas enriched with CS and/or HS necessary to verify their possible positive effects on newborns feeding in countries at high risk of infection and/or infestation.
Metabolites
Little is known about the extent of variation and activity of naturally occurring milk glycosidases and their potential to degrade milk glycans. A multi-omics approach was used to investigate the relationship between glycosidases and important bioactive compounds such as free oligosaccharides and O-linked glycans in bovine milk. Using 4-methylumbelliferone (4-MU) assays activities of eight indigenous glycosidases were determined, and by mass spectrometry and 1H NMR spectroscopy various substrates and metabolite products were quantified in a subset of milk samples from eight native North European cattle breeds. The results showed a clear variation in glycosidase activities among the native breeds. Interestingly, negative correlations between some glycosidases including β-galactosidase, N-acetyl-β-d-glucosaminidase, certain oligosaccharide isomers as well as O-linked glycans of κ-casein were revealed. Further, a positive correlation was found for free fucose content and α-fucosidase a...
Human milk glycosaminoglycans: the state of the art and future perspectives
Italian Journal of Pediatrics, 2013
Recently, a complete characterization and detailed evaluation of the glycosaminoglycans of human milk were performed. The total glycosaminoglycans content in milk from healthy mothers having delivered term or preterm newborns showed a constant pattern which was essentially composed of two main polysaccharides: chondroitin sulfate (60-70%) and heparin (30-40%). Moreover, considerable variations of glycosaminoglycans concentration were found during the first month of lactation, the highest values being present in colostrum compared to mature milk. Metabolism and potential biological functions of human milk glycosaminoglycans are hypothesized and future studies are encouraged.
Food Chemistry, 2019
Human milk oligosaccharides (HMOs) are free glycans naturally present in human milk that act as prebiotics, prevent pathogen binding, modulate the immune system and support brain development in infants. The HMOs composition and concentrations vary significantly among different women mainly because of the direct influence of the Secretor and Lewis phenotypes on HMOs biosynthesis. Analytical methods that can identify the differences in the HMOs composition and concentrations are a fundamental tool in HMOs research. This paper describes a simple HMOs extraction and analysis for the simultaneous and absolute quantification of neutral and acidic HMOs by graphitized carbon liquid chromatography-electrospray ionization-mass spectrometry. This method was validated and applied to analyze HMOs in the human milk obtained from 10 women. This method allows accurate and reliable quantification of HMOs and can be used to determine differences in HMOs concentrations throughout lactation and among women with different Secretor and Lewis phenotypes.
Milk Oligosaccharide Profiles by Reversed-Phase HPLC of Their Perbenzoylated Derivatives
Analytical Biochemistry, 1997
and pathogens involved in diseases of infants (2). These Human milk is rich in oligosaccharides, some of oligosaccharides are the third largest solid component in which inhibit toxins and pathogens involved in dismilk after lactose and triglycerides and are believed to eases of infants. To investigate qualitative and quantibe synthesized by the same glycosyltransferases that are tative individual variation of human milk oligosacchainvolved in the biosynthesis of glycoproteins and other rides, a sensitive method for routine identification and glycoconjugates. As a consequence, milk oligosaccharides quantification of intact milk oligosaccharides was decan have terminal carbohydrate structures similar to veloped and applied to milk samples from 50 donors. those present on glycoconjugates, some of which act as The isolated, reduced neutral oligosaccharide fraccell surface receptors for pathogens. Thus, some of these tions were perbenzoylated, resolved by reversedoligosaccharides could act as analogs or homologs to phase HPLC, and detected at 229 nm. This method rethose receptors, thereby providing protection to the nurssolves most structural isomers and does not require ing infant against enteric pathogens. Recent studies on stringent removal of lactose. Peaks were detected at complex sugar structures (3) have revealed the existence the low nanogram (pmol) level and peak areas were of a far greater number of oligosaccharides than what linear from 1 to 1000 mg for a standard oligosaccharide.
OMICS-rooted studies of milk proteins, oligosaccharides and lipids
Journal of Proteomics, 2009
Milk has co-evolved with mammals and mankind to nourish their offspring and is a biological fluid of unique complexity and richness. It contains all necessary nutrients for the growth and development of the newborn. Structure and function of biomolecules in milk such as the macronutrients (glyco-) proteins, lipids, and oligosaccharides are central topics in nutritional research. Omics disciplines such as proteomics, glycomics, glycoproteomics, and lipidomics enable comprehensive analysis of these biomolecule components in food science and industry. Mass spectrometry has largely expanded our knowledge on these milk bioactives as it enables identification, quantification and characterization of milk proteins, carbohydrates, and lipids. In this article, we describe the biological importance of milk macronutrients and review the application of proteomics, glycomics, glycoproteomics, and lipidomics to the analysis of milk. Proteomics is a central platform among the Omics tools that have more recently been adapted and applied to nutrition and health research in order to deliver biomarkers for health and comfort as well as to discover beneficial food bioactives.
A comparative study of free oligosaccharides in the milk of domestic animals
The British journal of nutrition, 2014
The present study was conducted to obtain a comprehensive overview of oligosaccharides present in the milk of a variety of important domestic animals including cows, goats, sheep, pigs, horses and dromedary camels. Using an analytical workflow that included ultra-performance liquid chromatography-hydrophilic interaction liquid chromatography with fluorescence detection coupled to quadrupole time-of-flight MS, detailed oligosaccharide libraries were established. The partial or full characterisation of the neutral/fucosylated, phosphorylated and sialylated structures was facilitated by sequencing with linkage- and sugar-specific exoglycosidases. Relative peak quantification of the 2-aminobenzamide-labelled oligosaccharides provided additional information. Milk from domestic animals contained a much larger variety of complex oligosaccharides than was previously assumed, and thirteen of these structures have been identified previously in human milk. The direct comparison of the oligosac...