Characterization and comparison of whey N-glycoproteomes from human and bovine colostrum and mature milk

•Whey N-glycoproteins were identified in human and bovine milk using glycoproteomics.•The bioinformatics analysis of the identified N-glycoproteins was conducted.•Immune-related N-glycoproteins were different in human and bovine milks.•Whey N-glycoproteomes varied between human and bovine milks across lactation stages. Milk glycoproteins are crucial nutrients with a variety of functions. However, whey N-glycoproteomes in human and bovine milks have not been characterized during lactation. Herein, using lectin enrichment and liquid chromatography tandem mass spectrometry, 68, 58, 100, and 98 N-glycoproteins were identified in human colostrum and mature milk as well as bovine colostrum and mature milk whey. Gene Ontology and KEGG pathway analyses were used to elucidate the biological functions of whey N-glycoproteins in human and bovine colostrum and mature milks. Whey N-glycoproteomes differed dramatically between human and bovine milks and across lactation stages. The conserved and specific whey N-glycoproteins in all four sample types were also determined. Our results improve understanding of the properties and biological functions of whey N-glycoproteins in human and bovine milk and colostra, and provide insight into the potential application of some N-glycoproteins in infant formulae at different stages of development.