Impact of Storage Conditions on the Breast Milk Peptidome

Howland, Vanessa; Klaedtke, Maik; Ruhnau, Johanna; Dhople, Vishnu M.; Grabe, Hans J.; Völker, Uwe; Heckmann, Matthias; Hammer, Elke

doi:10.3390/nu12092733

Cited by 12 publications

(11 citation statements)

References 41 publications

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“…Regardless of the samples being handled differently, the difficulties of human milk storage conditions in neonatal hospitals are widely recognized. Additionally, an investigation on the impact of storage conditions on the breast milk peptidome for MS experiments demonstrated that the majority of peptides kept quite stable when stored at −80°C, −20°C, or even at 4°C (Howland et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Quantification of FAM20A in human milk and identification of calcium metabolism proteins

Patel

Klootwijk

Whiting

et al. 2021

Physiological Reports

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Background FAM20A, a recently discovered protein, is thought to have a fundamental role in inhibiting ectopic calcification. Several studies have demonstrated that variants of FAM20A are causative for the rare autosomal recessive disorder, enamel‐renal syndrome (ERS). ERS is characterized by defective mineralization of dental enamel and nephrocalcinosis suggesting that FAM20A is an extracellular matrix protein, dysfunction of which causes calcification of the secretory epithelial tissues. FAM20A is a low‐abundant protein that is difficult to detect in biofluids such as blood, saliva, and urine. Thus, we speculated the abundance of FAM20A to be high in human milk, since the secretory epithelium of lactating mammary tissue is involved in the secretion of highly concentrated calcium. Therefore, the primary aim of this research is to describe the processes/methodology taken to quantify FAM20A in human milk and identify other proteins involved in calcium metabolism. Method This study used mass spectrometry‐driven quantitative proteomics: (1) to quantify FAM20A in human milk of three women and (2) to identify proteins associated with calcium regulation by bioinformatic analyses on whole and milk fat globule membrane fractions. Results Shotgun MS/MS driven proteomics identified FAM20A in whole milk, and subsequent analysis using targeted proteomics also successfully quantified FAM20A in all samples. Combination of sample preparation, fractionation, and LC‐MS/MS proteomics analysis generated 136 proteins previously undiscovered in human milk; 21 of these appear to be associated with calcium metabolism. Conclusion Using mass spectrometry‐driven proteomics, we successfully quantified FAM20A from transitional to mature milk and obtained a list of proteins involved in calcium metabolism. Furthermore, we show the value of using a combination of both shotgun and targeted driven proteomics for the identification of this low abundant protein in human milk.

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Section: Discussionmentioning

confidence: 99%

Quantification of FAM20A in human milk and identification of calcium metabolism proteins

Patel

Klootwijk

Whiting

et al. 2021

Physiological Reports

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“…The lowest lysozyme level in human milk was found in samples stored at room temperature. Howland et al (2020) reported cleavage in the peptide structure of the amino acid lysine, arginine, and glutamate due to an increased thrombin activity and other protease enzymes in human milk stored at room temperature. Lysozyme or muramidase is a part of group of glycosidic hydrolases with 14.7 kDa of atomic mass in human lysozyme.…”

Section: Discussionmentioning

confidence: 99%

Stability of lactoferrin and lysozyme in human milk at various temperatures and duration of storage

Kasiati¹,

Pertami²,

Utomo³

et al. 2021

Mal J Nutr

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Introduction: Exclusive breastfeeding, especially in the first six months, is essential for infants as it provides nutrition and protection against various diseases. Colostrum, which is found in the first breast milk produced, contains various protective factors, such as lactoferrin and lysozyme. Human milk can be stored at room temperature, refrigerated, or kept frozen. Several factors affect the stability of the bioactive content in human milk, such as temperature and storage time. The aim of this study was to measure the stability of lactoferrin and lysozyme levels in human milk during the first six hours (h) at different temperatures and compare it with that of frozen human milk. Methods: Human milk samples were obtained from 11 breastfeeding mothers using certain criteria. The human milk was stored at room temperature and 4°C for 1, 3, and 6 h and classified as never frozen, while frozen human milk was stored at -20°C for 1, 3, and 6 days. Measurement of the lactoferrin and lysozyme levels was performed using enzyme-linked immunosorbent assay. Results: The results showed that storage at room temperature significantly reduced lactoferrin and lysozyme levels. Lactoferrin levels in frozen human milk did not significantly decrease during the first six days. Meanwhile, the lysozyme levels in frozen human milk decreased significantly. Conclusion: The levels of lactoferrin and lysozyme in frozen human milk stored for the first six days were more stable than those stored at room temperature and 4°C in the first 6 h.

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“…Additionally, human milk and feces need to be collected weekly and stored in the correct way to allow for microbiota analysis. Storage conditions, including temperature, have been shown to influence human milk peptidome and fecal microbiota composition [65][66][67][68]. Lastly, the absorption of proteins from human milk in the small intestine cannot be measured directly, although it may influence metabolic activity of the microbiota in the colon.…”

Section: Discussionmentioning

confidence: 99%

From Mum to Bum: An Observational Study Protocol to Follow Digestion of Human Milk Oligosaccharides and Glycoproteins from Mother to Preterm Infant

et al. 2021

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The nutritional requirements of preterm infants are challenging to meet in neonatal care, yet crucial for their growth, development and health. Aberrant maturation of the gastrointestinal tract and the microbiota could affect the digestion of human milk and its nutritional value considerably. Therefore, the main objective of the proposed research is to investigate how the intestinal microbiota of preterm and full-term infants differ in their ability to extract energy and nutrients from oligosaccharides and glycoproteins in human milk. This pilot study will be an observational, single-center study performed at the Neonatal Intensive Care Unit at Isala Women and Children’s Hospital (Zwolle, The Netherlands). A cohort of thirty mother–infant pairs (preterm ≤30 weeks of gestation, n = 15; full-term 37–42 weeks of gestation, n = 15) will be followed during the first six postnatal weeks with follow-up at three- and six-months postnatal age. We will collect human milk of all mothers, gastric aspirates of preterm infants and fecal samples of all infants. A combination of 16S rRNA amplicon sequencing, proteomics, peptidomics, carbohydrate analysis and calorimetric measurements will be performed. The role of the microbiota in infant growth and development is often overlooked yet offers opportunities to advance neonatal care. The ‘From Mum to Bum’ study is the first study in which the effect of a preterm gut microbiota composition on its metabolic capacity and subsequent infant growth and development is investigated. By collecting human milk of all mothers, gastric aspirates of preterm infants and fecal samples of all infants at each timepoint, we can follow digestion of human milk from the breast of the mother throughout the gastrointestinal tract of the infant, or ‘From Mum to Bum’.

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Impact of Storage Conditions on the Breast Milk Peptidome

Cited by 12 publications

References 41 publications

Quantification of FAM20A in human milk and identification of calcium metabolism proteins

Quantification of FAM20A in human milk and identification of calcium metabolism proteins

Stability of lactoferrin and lysozyme in human milk at various temperatures and duration of storage

From Mum to Bum: An Observational Study Protocol to Follow Digestion of Human Milk Oligosaccharides and Glycoproteins from Mother to Preterm Infant

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