Antibody-independent identification of bovine milk-derived peptides in breast-milk

Picariello, Gianluca; Addeo, Francesco; Ferranti, Pasquale; Nocerino, Rita; Paparo, Lorella; Passariello, Annalisa; Dallas, David C.; Robinson, Randall C.; Canani, Roberto Berni

doi:10.1039/c6fo00731g

Cited by 14 publications

(25 citation statements)

References 44 publications

(59 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On average, more than 1,200 peptides derived from endogenous proteins were identified by HPLC-MS/MS in the 12% TCA-soluble peptide fraction of each breast milk sample (not shown), as already reported in our previous work ( 24 ). However, the repertory of the endogenous peptides exhibited a significant intra-individual variability, as clearly observed when comparing the chromatographic profiles.…”

Section: Resultssupporting

confidence: 85%

“…For the first time, we exploited MS to detect peptides arising from the digestion of bovine casein and whey proteins excreted into the breast milk of lactating donors after an oral administration of cow's milk ( 24 ). In particular, bovine β-Lg and α s1 -casein fragments were detected in the 12% trichloroacetic (TCA)-soluble fraction of two and one out six breast milk samples, respectively.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Excretion of Dietary Cow's Milk Derived Peptides Into Breast Milk

et al. 2019

Self Cite

View full text Add to dashboard Cite

Nanoflow-HPLC-tandem mass spectrometry (MS/MS) was used to analyze the peptide fraction of breast milk samples collected from a single non-atopic donor on different days (10 samples) after receiving an oral load of cow's milk (by drinking 200 mL of bovine milk). In addition, breast milk was sampled from the same lactating mother over a 6-h period at five time points after drinking cow's milk. We aimed to trace the intra-individual variability and to define a time profile of the excretion of dietary peptides into breast milk. Overall, 21 peptides exclusively originating from both bovine caseins and whey proteins with no match within the human milk proteome were identified in the breast milk samples. These peptides were missing in the breast milk obtained from the mother after a prolonged milk- and dairy-free diet (three samples). The time course of cow's milk-derived β-Lg f(125–135) and β-casein f(81–92) in breast milk was determined from the MS ion intensity of the peptide signals. No intact cow's milk gene products were detected by HPLC-MS/MS analysis and Western blotting with anti-β-Lg antibody, but dot-blot analysis confirmed the occurrence of β-Lg fragments in the enriched peptide fraction of breast milk. These data suggest shifting the analytical perspective for the detection of dietary food allergens in breast milk from intact proteins to digested peptide fragments. The possible sensitization and elicitation potential or the tolerogenic properties of such low amounts of dietary peptides for the breastfed newborns remain to be explored.

show abstract

Section: Resultssupporting

confidence: 85%

Section: Introductionmentioning

confidence: 99%

Excretion of Dietary Cow's Milk Derived Peptides Into Breast Milk

et al. 2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…Our study provides strong evidence for the presence of intact nonhuman proteins in human milk, from mothers not exposed to any dietary intervention. Notably, there was an earlier study 23 wherein they identiified the YVEELKPTPEGDL peptide from bovine Beta-lactoglobulin in human milk. We additionally searched raw files of previously published human milk proteomics studies deposited in PRIDE 38 (PXD00365 and PXD00296) using our search parameters and identified quite a few nonhuman tryptic peptides, mostly originating from bovine milk proteins (e.g., “HIQKEDVPSER” and “FFVAPFPEVFGK” from bovine Alpha-S1-casein, “YLGYLEQLLR” from bovine or sheep Alpha-S1-casein, “AVPYPQR” from bovine Beta-casein, “VLPVPQK” from bovine or sheep Beta-casein, “YIPIQYVLSR” from bovine or sheep Kappa-casein) that were also identified in the present study.…”

Section: Discussionmentioning

confidence: 99%

Discovery and Quantification of Nonhuman Proteins in Human Milk

et al. 2018

View full text Add to dashboard Cite

The question whether and which nonhuman peptides or proteins are present in human milk was raised many decades ago. However, due to cross-reactivity or nonspecific antibody recognition, the accuracy of detection by immunochemical methods has been a concern. Additionally, the relative low-abundance of nonhuman peptides/proteins in the complex milk sample makes them a challenging target to detect. Here, by deep proteome profiling, we detected several nonhuman peptides, which could be grouped as nonhuman proteins. We next estimated their concentration in human milk by combining data-dependent shotgun proteomics and parallel reaction monitoring. First, we fractionated human milk at the protein level and were able to detect 1577 human proteins. Additionally, we identified 109 nonhuman peptides, of which 71 were grouped into 36 nonhuman proteins. In the next step, we targeted 37 nonhuman peptides and nine of them could be repeatedly quantified in human milk samples. Peptides/proteins originating from bovine milk products were the dominant nonhuman proteins observed, notably bovine caseins (α-S1-, α-S2-, β-, κ-caseins) and β-lactoglobulin. The method we present here can be expanded to investigate more about nonhuman peptides and proteins in human milk and give a better understanding of how human milk plays a role in allergy prevention.

show abstract

“…The relatively high levels of BLG peptide sequences that we found in milk from allergic mothers explains the high sequence coverage of BLG in the current study when compared to Zhu et al [2]. Because we removed small peptides by filter-aided sample preparation (10-20 kDa cutoff), no comparison could be made with previous peptidomics studies [3,4]. Other qualitative differences with these previous studies can be attributed to the more strict filtering on false positives that we applied, the inclusion of serum albumins, the inclusion of semi-trypsin and non-trypsin-digested sequences, and to the inclusion of milk from allergic mothers.…”

Section: Discussionmentioning

confidence: 52%

“…According to studies using mass spectrometry, the main biological source of the nonhuman proteinaceous molecules in human milk seems to be bovine milk. A peptidomics study demonstrated the presence of two peptides originating from bovine β-lactoglobulin (BLG) and one originating from α S1 -casein [3]. In a later study, this was extended with peptides from α-lactalbumin (ALA), κ-casein, β-casein, and lactoferrin [4].…”

Section: Introductionmentioning

confidence: 97%

Maternal Allergy and the Presence of Nonhuman Proteinaceous Molecules in Human Milk

et al. 2020

View full text Add to dashboard Cite

Human milk contains proteins and/or protein fragments that originate from nonhuman organisms. These proteinaceous molecules, of which the secretion might be related to the mother’s allergy status, could be involved in the development of the immune system of the infant. This may lead, for example, to sensitization or the induction of allergen-specific tolerance. The aim of this study was to investigate the relation between maternal allergy and the levels of nonhuman proteinaceous molecules in their milk. In this study, we analysed trypsin-digested human milk serum proteins of 10 allergic mothers and 10 nonallergic mothers. A search was carried out to identify peptide sequences originating from bovine or other allergenic proteins. Several methods were applied to confirm the identification of these sequences, and the differences between both groups were investigated. Out of the 78 identified nonhuman peptide sequences, 62 sequences matched Bos taurus proteins. Eight peptide sequences of bovine β -lactoglobulin had significantly higher levels in milk from allergic mothers than in milk from nonallergic mothers. Dietary bovine β -lactoglobulin may be absorbed through the intestinal barrier and secreted into human milk. This seems to be significantly higher in allergic mothers and might have consequences for the development of the immune system of their breastfed infant.

show abstract

Antibody-independent identification of bovine milk-derived peptides in breast-milk

Cited by 14 publications

References 44 publications

Excretion of Dietary Cow's Milk Derived Peptides Into Breast Milk

Excretion of Dietary Cow's Milk Derived Peptides Into Breast Milk

Discovery and Quantification of Nonhuman Proteins in Human Milk

Maternal Allergy and the Presence of Nonhuman Proteinaceous Molecules in Human Milk

Contact Info

Product

Resources

About