A novel strategy for subtracting high-abundance proteins in human milk by immunoaffinity chromatography / 解放军医学杂志

ABSTRACT

Objective To establish and evaluate a novel method for subtracting high-abundance proteins in order to enrich low abundance proteins in human milk, with the aim of discovering important bioactive factors by proteomics approach in the future. Methods The whole proteins in human milk were used as immunogens to prepare polyclonal antibody, and the high-abundance proteins were subtracted by immunoaffinity chromatography. The effect was subsequently evaluated by immunoblotting. Results Anti-human milk polyclonal antibody was successfully prepared following routine method. After immunoaffinity chromatography analysis, four kinds of high-abundance proteins in human milk were removed. Conclusions These data indicated that the anti-human milk polyclonal antibodies prepared by whole protein samples were available for the removal of high-abundance proteins and enriching low abundance proteins of the samples. Accordingly, this method could also be used in detecting low abundance proteins in sample such as serum.