Comprehensive proteomic analysis of the human milk proteome: contribution of protein fractionation.
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.
Mangé
A
A
University of Montpellier I, Montpellier, France.
Bellet
V
V
Tuaillon
E
E
Van de Perre
P
P
Solassol
J
J
D005591:Chemical Fractionation / Q000379:methods*; D005260:Female; D006801:Humans; D008895:Milk, Human / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D011506:Proteins; D020543:Proteome