Sixteen hybridoma-derived antibodies to the epidermal growth factor receptor of A431 ceils were studied with respect to their reactions with blood group-related carbohydrate structures. Twelve of these were assessed as recognizing carbohydrate determinants on the basis of their immunostaining of reference blood group substances on nitrocellulose paper. Three of these antibodies were further investigated by inhibition of binding assays with giycoproteins and structurally defined oligosaccharides or by haemagglutination of erythrocytes before and after treatment with endo-β-galactosidase. Two of the antibodies, 29.1 and 455, were shown to have blood group A-related specificities which differed from one another and from those of monocional anti-A antibodies described previously. The third antibody, 3CIB12, which was shown to recognize a determinant based on αl+3 fucosylated Type 2 chains on linear and branched backbone sequences, also differs from previously described monoclonal antibodies of 3-fucosyl-N-acetyllactosamine type, such as anti-SSEA-1 (anti-mouse embryo) and several antibodies to human myeloid ceils. While these antibodies are invaluable in providing structural information on the carbohydrate chains of the receptor glycoprotein and should help to elucidate their functions, their use as ‘anti-receptor’ reagents in cell biology will be influenced by the knowledge that the determinants they recognize are shared by other glycoproteins and glycolipids of diverse cell types.
3P172 A Quantitative Analysis of Epidermal Growth Factor Receptor Clustering Using Super-resolution Microscopy(Cell biology,Poster,The 52th Annual Meeting of the Biophysical Society of Japan(BSJ2014))
