Identification of carbohydrate deficient transferrin forms by MALDI-TOF mass spectrometry and lectin ELISABiochim...

Transferrin was isolated from sera of patients with severe alcohol abuse and from control sera by affinity chromatography using an immobilized polyclonal antibody from sheep, followed by gel filtration. The purified transferrin was then separated by MonoQ chromatography. Compared to the controls, sera from heavy alcohol consumers showed two additional transferrin peaks, eluting earlier than the three main transferrin forms present in all sera. Further analysis of the isolated transferrin forms by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and enzyme linked immunosorbent assay with different digoxigenylated lectins (lectin ELISA) revealed that the main carbohydrate deficient transferrin (CDT) forms are lacking either one or both of the N-Glycan chains.