Offline Coupling of Low-Pressure Anion-Exchange Chromatography with MALDI-MS to Determine the Elution Order of Human Milk Oligosaccharides

2000 ◽  
Vol 284 (2) ◽  
pp. 256-265 ◽  
Author(s):  
Berndt Finke ◽  
Marko Mank ◽  
Hannelore Daniel ◽  
Bernd Stahl
Keyword(s):  
Human Milk ◽  
Anion Exchange ◽  
Low Pressure ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Maldi Ms ◽  
Elution Order ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides

scholarly journals Systematic Characterization and Longitudinal Study Reveal Distinguishing Features of Human Milk Oligosaccharides in China

2020 ◽  
Vol 4 (8) ◽  
Author(s):  
Jiayi Wu ◽  
Shaohui Wu ◽  
Jinhong Huo ◽  
Hongbo Ruan ◽  
Xiaofei Xu ◽  
...  
Keyword(s):  
Longitudinal Study ◽  
Human Milk ◽  
High Performance ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
African Populations ◽  
Chinese Mothers ◽  
Distinguishing Features

ABSTRACT Background Human milk oligosaccharides (HMOs) in breast milk contribute to the development of the neonatal microbiota and immune system. However, longitudinal studies examining HMO profiles of Chinese mothers remain scarce. Objectives We aimed to analyze HMO profiles, including their composition, concentrations, and changes during lactation, in milk of Chinese mothers. Methods A total of 822 milk samples from 222 mothers were collected, of which 163 mothers provided single samples. Samples from the remaining 59 mothers were collected on day 3, day 7, and thereafter every 7 or 14 d until day 168. 24 HMOs were studied using high-performance anion-exchange chromatography. Secretor and nonsecretor status were determined based on Lewis blood types and a defined 2′-fucosyllactose (2′-FL) threshold. Results Of the 222 mothers, 77% were secretors and 23% were nonsecretors. The longitudinal study involving 59 mothers showed that the total HMOs in secretors were significantly greater than those in nonsecretors during the first 2 wk. Acidic HMOs decreased significantly during lactation and were similar between secretors and nonsecretors. Among neutral HMOs, distinctive differences were observed. Nonfucosylated and α-1-3/4-fucosylated HMOs in nonsecretors were significantly higher than those in secretors during the first month. In contrast, α-1-2-fucosylated HMOs in secretors were significantly higher than those in nonsecretors throughout 168 d. In secretors, 2′-FL concentrations peaked at (mean ± SEM) 3.02 ± 0.14 g/L (day 3) followed by significant decreases. In nonsecretors, 2′-FL concentrations were fairly low throughout 168 d. Of the 24 studied HMOs, only 3-fucosyllactose concentrations increased during lactation in both secretor and nonsecretor mothers. Conclusions Our study showed dynamic changes of 24 HMOs in secretors and nonsecretors during lactation and revealed unique features of these HMO profiles in the milk of Chinese mothers. Interestingly, 2′-FL concentrations in secretors were found to be lower than those of Western populations but higher than those of African populations.

scholarly journals Neutral oligosaccharide content of preterm human milk

1999 ◽  
Vol 82 (5) ◽  
pp. 361-367 ◽  
Author(s):  
Tarek Nakhla ◽  
Daotian Fu ◽  
David Zopf ◽  
Nancy L. Brodsky ◽  
Hallam Hurt
Keyword(s):  
Human Milk ◽  
Gestational Age ◽  
High Performance ◽  
Gel Filtration ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Milk Samples ◽  
Neutral Oligosaccharide

Human milk oligosaccharides are known to play a role in protection against certain infectious diseases. Previous reports indicate that the content of human milk oligosaccharides varies widely among individuals at term but such information on preterm milk is lacking. After removal of the fat, protein and most of the lactose from non-pooled human milk samples, a total neutral oligosaccharide fraction was isolated by ion-exchange chromatography followed by gel filtration. A Dionex high-performance anion-exchange chromatography system equipped with a pulsed electrometric detector was then employed to measure the levels of ten neutral oligosaccharides in the individual milk samples. Twenty-three milk samples from thirteen mothers who delivered at a mean gestational age of 29·5 (sd 3·1) weeks were collected between days 0 and 33 of lactation, and compared with three samples of term milk from two mothers. The ranges of the total and individual levels of the ten neutral oligosaccharides in preterm milk were similar to those in term milk. Further, as previously described in term milk, preterm milk exhibited a quantitative individual variation. This variation was independent of the gestational age, day of lactation, and postconceptional age. In conclusion, levels of ten neutral oligosaccharides did not differ between preterm and term human milk.

scholarly journals Variation of human milk oligosaccharides in relation to milk groups and lactational periods

2010 ◽  
Vol 104 (9) ◽  
pp. 1261-1271 ◽  
Author(s):  
Stephan Thurl ◽  
Manfred Munzert ◽  
Jobst Henker ◽  
Günther Boehm ◽  
Beate Müller-Werner ◽  
...  
Keyword(s):  
Human Milk ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Blood Type ◽  
Human Milk Oligosaccharides ◽  
Milk Oligosaccharides ◽  
Milk Samples ◽  
Test Kit ◽  
Biological Protection ◽  
Group 2

Human milk oligosaccharides, representing the third largest fraction of human milk, have been assigned important protective functions for newborns acting as bifidogenic substrates or as inhibitory agents towards pathogens. Using high-pH anion-exchange chromatography and an enzyme test kit, twenty oligosaccharides and lactose were determined in milk samples of German women from days 3 to 90 postpartum. Twenty-two secretor mothers with Lewis blood group Le(a − b+) synthesised all twenty oligosaccharides, and could be assigned to milk group 1. Five non-secretor mothers (Le(a+b − )) produced all oligosaccharides with the exception of α1,2-fucosylated compounds (milk group 2), whereas three secretor mothers with blood type Le(a − b − ) lacked α1,4-fucosyloligosaccharides, corresponding to milk group 3. Secretor women of milk groups 1 and 3 synthesised significantly higher amounts of total neutral oligosaccharides and of several total core structures (e.g. lacto-N-tetraose) than non-secretor women. Generally, these oligosaccharides significantly decrease during the first 3 months postpartum. By comparing fucosyloligosaccharides within and among the three milk groups, insight into their biosynthesis could be gained. Six acidic oligosaccharides without fucose residues were detected in milk samples of all mothers. Regression analysis confirmed that total acidic oligosaccharides declined threefold during the study period. Milk samples corresponding to the three milk groups exhibited significant qualitative and quantitative differences during the first 3 months of lactation. It can be assumed that particularly milk of non-secretor women (milk group 2) exerts a modified biological protection in the babies in comparison with milks of secretors (groups 1 and 3).

scholarly journals Low Diversity of Human Milk Oligosaccharides is Associated with Necrotising Enterocolitis in Extremely Low Birth Weight Infants

Nutrients ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 1556 ◽  
Author(s):  
Erik Wejryd ◽  
Magalí Martí ◽  
Giovanna Marchini ◽  
Anna Werme ◽  
Baldvin Jonsson ◽  
...  
Keyword(s):  
Birth Weight ◽  
Breast Milk ◽  
Low Birth Weight ◽  
Human Milk ◽  
Amperometric Detection ◽  
Exchange Chromatography ◽  
Extremely Low Birth Weight ◽  
Human Milk Oligosaccharides ◽  
Low Birth Weight Infants ◽  
Milk Oligosaccharides

Difference in human milk oligosaccharides (HMO) composition in breast milk may be one explanation why some preterm infants develop necrotizing enterocolitis (NEC) despite being fed exclusively with breast milk. The aim of this study was to measure the concentration of 15 dominant HMOs in breast milk during the neonatal period and investigate how their levels correlated to NEC, sepsis, and growth in extremely low birth weight (ELBW; <1000 g) infants who were exclusively fed with breast milk. Milk was collected from 91 mothers to 106 infants at 14 and 28 days and at postmenstrual week 36. The HMOs were analysed with high-performance anion-exchange chromatography with pulsed amperometric detection. The HMOs diversity and the levels of Lacto-N-difucohexaose I were lower in samples from mothers to NEC cases, as compared to non-NEC cases at all sampling time points. Lacto-N-difucohexaose I is only produced by secretor and Lewis positive mothers. There were also significant but inconsistent associations between 3′-sialyllactose and 6′-sialyllactose and culture-proven sepsis and significant, but weak correlations between several HMOs and growth rate. Our results suggest that the variation in HMO composition in breast milk may be an important factor explaining why exclusively breast milk fed ELBW infants develop NEC.

scholarly journals Structural composition and functional characterization of soluble CD59: heterogeneity of the oligosaccharide and glycophosphoinositol (GPI) anchor revealed by laser-desorption mass spectrometric analysis

1996 ◽  
Vol 316 (3) ◽  
pp. 923-935 ◽  
Author(s):  
Seppo MERI ◽  
Timo LEHTO ◽  
Chris W. SUTTON ◽  
Jaana TYYNELÄ ◽  
Marc BAUMANN
Keyword(s):  
Anion Exchange ◽  
Functional Characterization ◽  
Laser Desorption ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Maldi Ms ◽  
Spectrometric Analysis ◽  
Gpi Anchor ◽  
Laser Desorption Mass Spectrometry ◽  
Structural Composition

CD59 (protectin) is a glycophosphoinositol (GPI)-anchored inhibitor of the membrane attack complex of complement found on blood cells, endothelia and epithelial cells. In addition to the lipid-tailed CD59, soluble lipid-free forms of CD59 are present in human body fluids. We have investigated the detailed structural composition of the naturally occurring soluble urinary CD59 (CD59U) using peptide mapping, anion-exchange chromatography, sequential exoglycosidase digestion and matrix-assisted laser-desorption mass spectrometry (MALDI-MS). CD59U exhibited an average Mr of 12444 in MALDI-MS. Mass analysis of the isolated C-terminal peptide (T9) indicated that a GPI-anchor (at Asn-77) without an inositol-associated phospholipid was present in soluble CD59U. By using residue-specific exoglycosidases, chemical modification and MALDI-MS structures of seven different GPI-anchor variants were determined. Variant forms of the anchor had deletions and/or extensions of one or more monosaccharide units. Sialic acid linked to an N-acetylhexosamine-galactose arm was found in two GPI-anchor variants. The N-linked carbohydrate side chain of CD59U (at Asn-18) also displayed considerable heterogeneity. The predominant oligosaccharide chains were fucosylated biantennary and triantennary complexes with variable sialylation. Mono Q anion-exchange chromatography resolved urinary CD59 into nine different fractions that bound equally well to the terminal complement SC5b–8 complexes. Despite binding to C5b–8, soluble CD59U inhibited complement lysis at an approx. 200-fold lower efficiency than erythrocyte CD59. These results document the structural heterogeneity of both the GPI anchor and N-linked oligosaccharide of CD59 and demonstrate that the phospholipid tail is needed for the full functional activity of CD59. The site of cleavage between the diradylglycerol phosphate and inositol suggests that a mammalian phospholipase D could be involved in the solubilization of GPI-anchored proteins.

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