alexa Comprehensive proteomic analysis of the human milk proteome: contribution of protein fractionation.


Biosensors Journal

Author(s): Mang A, Bellet V, Tuaillon E, Van de Perre P, Solassol J, Mang A, Bellet V, Tuaillon E, Van de Perre P, Solassol J

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Abstract In-depth analysis of the milk proteome by mass spectrometry is challenged by the presence of few high-abundance proteins that interfere with the detection of lower-abundance proteins. Here, we evaluated the proteomic analysis of milk samples following a strong anion exchange fractionation procedure using denaturating conditions ensuring the disruption of protein-protein interactions. Crude whey or skim milk and their different resulting fractions were analyzed by protein chip array mass spectrometry. Using protein chip array mass spectrometry, several high-abundance proteins were localized in distinct fractions increasing the total number of unique peptides and proteins detected. This total number increased by about 20-30\% by combining different chromatographic surface arrays used for capture. Reproducible results were obtained in human skim milk and whey; however this approach was not successful with milk fat globule membrane and required refinement. Hence, milk profiling by anion exchange fractionation combined to protein chip array mass spectrometry represents a promising tool to detect unknown low-abundance milk proteins that may ultimately prove useful as biomarkers of diseases transmitted by breastfeeding. This article was published in J Chromatogr B Analyt Technol Biomed Life Sci and referenced in Biosensors Journal

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