Characterization of the binding epitope of a monoclonal antibody to sulphatide.
An IgG1 monoclonal antibody, Sulph I, reacting with sulphatide (3'-sulphogalactosylceramide), was produced by immunizing Balb/c mice with that glycolipid coated on Salmonella minnesota bacterial membrane. Radioimmunodetection of the binding of the monoclonal antibody to structurally related glycolipids adsorbed to microtitre plates or chromatographed on thin-layer plates was used to determine its binding epitope. The antibody showed similar binding avidity to three sulphated glycolipids: sulphatide, sulpholactosylceramide and seminolipid. Lysosulphatide did bind the antibody, but, compared with sulphatide, 30 times more antigen was needed for half-maximal binding. Bis(sulphogangliotriosyl)ceramide and bis-sulphogangliotetraosylceramide did not bind the antibody. These results suggest that terminal galactose-3-O-sulphate and part of the hydrophobic region of the glycolipid are recognized by the Sulph I antibody.
Fredman
P
P
Department of Psychiatry and Neurochemistry, University of Gothenburg, Sweden.
Mattsson
L
L
Andersson
K
K
Davidsson
P
P
Ishizuka
I
I
Jeansson
S
S
Månsson
J E
JE
Svennerholm
L
L
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002554:Cerebrosides / Q000276:immunology*; D002855:Chromatography, Thin Layer; D000939:Epitopes / Q000276:immunology*; D005699:Galactosylceramides / Q000276:immunology*; D006017:Glycolipids / Q000276:immunology; D011863:Radioimmunoassay; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D002554:Cerebrosides; D000939:Epitopes; D005699:Galactosylceramides; D006017:Glycolipids