It was previously observed that changes in the expression of specific proteins as well as protein glycosylation occur with differentiation. In this study, specific mucin O-glycans were identified that mark butyrate-induced epithelial differentiation of the intestinal cell line CaCo-2, by applying porous graphitized carbon nano-liquid chromatography with electrospray ionization tandem mass spectrometry (PGC-nanoLC-MS/MS). Moreover, a quantitative proteomic approach was used to decipher changes in the cell proteome. By performing an integrative approach, we generated hypotheses about glycosylation signatures of specific cell adhesion proteins.