This study applied yolk
immunoglobulins immunoaffinity separation and MALDI-TOF MS for clinical proteomics of
congenital disorders of glycosylation (CDG) and secondary glycosylation disorders [
galactosemia and
hereditary fructose intolerance (HFI)]. Serum
transferrin (Tf) and alpha1-antitrypsin (AAT) that are markers for CDG, were purified sequentially to obtain high-quality MALDI mass spectra to differentiate single glycoforms of the native intact
glycoproteins. The procedure was found feasible for the investigation of
protein macroheterogeneity due to glycosylation site underoccupancy then ensuing the characterization of patients with CDG group I (N-
glycan assembly disorders). Following
PNGase F digestion of the purified
glycoprotein, the characterization of
protein microheterogeneity by N-
glycan MS analysis was performed in a patient with CDG group II (processing disorders). CDG-Ia patients showed a typical profile of underglycosylation where the fully glycosylated glycoforms are always the most abundant present in plasma with lesser amounts of partially and unglycosylated glycoforms in this order.
Galactosemia and HFI are potentially fatal diseases, which benefit from early diagnosis and prompt therapeutic intervention. In symptomatic patients with
galactosemia and in those with HFI, MALDI MS of Tf and AAT depicts a hypoglycosylation profile with a significant increase of underglycosylated glycoforms that reverses by dietary treatment, representing a clue for diagnosis and treatment monitoring.