Monoclonal antibody against a lactose epitope of glycosphingolipids binds to melanoma tumour cells.
Mice were immunized with a neoglycoprotein consisting of a chemically modified carbohydrate moiety (reductively aminated 3'-sialyllactose) linked to human serum albumin. By this procedure an antibody response to the normally non-immunogenic carbohydrate structure was obtained. Hybridomas were established, and monoclonal antibodies were selected in ELISA based on their binding to the saccharide hapten, or to a lactosylceramide-mimicking neoglycolipid, lactose-bis-sulfone. One of the selected antibodies, 2H4, was of particular interest, since it also bound to glycolipids present on melanoma cells. FACS analysis of a panel of 14 melanoma cell lines showed that the 2H4 antibody bound to the majority of these. In frozen, non-fixed sections or paraffin sections of biopsies the monoclonal antibody 2H4 stained melanoma cells, but not tumour infiltrating lymphocytes or normal skin. Detailed immunochemical analysis of 2H4, using thin layer chromatography revealed that it recognized an internal lactose epitope in several glycosphingolipids.
Ding
K
K
Department of Tumor Immunology, Wallenberg Laboratory, University of Lund, Sweden.
Ekberg
T
T
Zeuthen
J
J
Teneberg
S
S
Karlsson
K A
KA
Rosén
A
A
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002240:Carbohydrate Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006028:Glycosphingolipids / Q000276:immunology*; D007785:Lactose / Q000276:immunology*; D008545:Melanoma / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D014407:Tumor Cells, Cultured; D014614:Vaccines, Synthetic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006028:Glycosphingolipids; D014614:Vaccines, Synthetic; D007785:Lactose