A murine monoclonal antibody detecting N-acetyl- and N-glycolyl-GM2: characterization of cell surface reactivity.
Antisera reactive with the ganglioside GM2 were raised by immunizing C57BL/6 mice with the C57BL/6 melanoma JB-RH. Fusion with NS-1 was performed using splenic mononuclear cells from a mouse with high antibody titer. An immunoglobulin M monoclonal antibody (monoclonal antibody 5-3) was identified which was reactive with an antigen that was resistant to heat, trypsin, and Pronase. A panel of purified glycolipids was used to determine the specificity of monoclonal antibody 5-3. Reactivity was restricted to N-acetyl- and N-glycolyl-GM2. No reactivity was detected with asialo-GM2 or other gangliosides. Monoclonal antibody 5-3 was used to define the expression of GM2 on the cell surface of cultured human normal and malignant cells. Reactivity was seen with cell lines derived from 8 of 8 astrocytomas, 5 of 5 neuroblastomas, 7 of 9 sarcomas, 4 of 18 human melanomas, 2 of 4 murine melanomas, 4 of 37 epithelial cancers and with 0 of 6 skin fibroblast and 0 of 2 brain fibroblast lines. GM2, like GD2 and GD3, appears to be a differentiation antigen largely restricted to cells of neuroectodermal origin.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer; D004797:Enzyme-Linked Immunosorbent Assay; D005678:G(M2) Ganglioside / Q000032:analysis*; D005732:Gangliosides / Q000032:analysis*; D051379:Mice; D008810:Mice, Inbred C57BL
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005732:Gangliosides; D005678:G(M2) Ganglioside