Improving Fractionation of Human Milk Proteins through Calcium Phosphate Coprecipitation and Their Rapid Characterization by Capillary Electrophoresis

Journal of Proteome Research
Isabel Ten-DoménechJosé Manuel Herrero-Martínez

Abstract

This work describes a simple sample pretreatment method for the fractionation of human milk proteins (into their two main groups, whey and caseins) prior to their analysis. The protein-extraction protocol is based on the addition of calcium phosphate to nonadjusted pH human milk. The combination of calcium ions with phosphate results in an effective coprecipitation of caseins. To assess the suitability of this fractionation protocol, the protein extracts were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), LC-MS/MS, and capillary electrophoresis (CE) analysis. The results evidence a significant decrease in contamination of casein fraction with whey proteins and vice versa compared with the conventional isoelectric precipitation of caseins. In addition, CE fraction collection coupled to LC-MS/MS (off-line coupling) has been successfully applied to the identification of minor proteins in this complex matrix. The methodology presented here constitutes a promising tool to enlarge the knowledge of human milk proteome.

References

Oct 1, 1974·Journal of Dairy Science·T NagasawaM Takase
Nov 1, 1994·Archives of Biochemistry and Biophysics·B C DevC W Slattery
Dec 10, 1996·Proceedings of the National Academy of Sciences of the United States of America·A ShevchenkoM Mann
Jun 2, 2001·Biotechnology Progress·T MorçölS J Bell
Aug 16, 2003·European Journal of Biochemistry·Esben S SørensenLone K Rasmussen
May 7, 2010·Journal of Proteome Research·Angelo D'AlessandroLello Zolla
Mar 3, 2011·Journal of Proteome Research·Yalin LiaoBo Lönnerdal
Feb 9, 2012·Journal of Proteome Research·Claire E MolinariPeter E Hartmann
Mar 6, 2012·Annual Review of Food Science and Technology·Douglas G Dalgleish, Milena Corredig
Jul 10, 2012·Analytical and Bioanalytical Chemistry·Munazza Raza MirzaGünther K Bonn
Mar 6, 2013·Journal of Paediatrics and Child Health·Bo Lönnerdal
Jan 1, 2014·Preparative Biochemistry & Biotechnology·Chon-Ho YenChing-Fu Tu
Apr 15, 2014·Nature Biotechnology·Juan A VizcaínoHenning Hermjakob
Oct 23, 2016·Biology Direct·Lukasz P Kozlowski

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Citations

Jan 31, 2020·Expert Review of Proteomics·Esei T TeclemariamStephanie M Cologna

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