Chemical Identification of Glycosphingolipid Antigens Recognized by Antibodies.

A number of monoclonal antibodies have been generated aiming to detect markers for tissue development and degeneration, cell differentiation, and malignant transformation. Among antigens recognized by these monoclonal antibodies, glycosphingolipids including gangliosides are included. In this chapter, fundamental methods for chemical identification of ganglioside antigens recognized by antibodies are summarized. After extraction of lipids from cells and tissues using organic solvent, neutral and acidic glycosphingolipids are separated with/without pre-elimination of phospholipids. Extracted glycolipids are applied for thin-layer chromatography combined with immunodetection to identify specific antigen component recognized by the antibody. To finally define the antigen structure, mass spectrometry is effectively used. Confirmation with transfection of supposed glycosyltransferase cDNA has also been a supportive way.