Generation and characterization of the binding epitope of a novel monoclonal antibody to sulfatide (sulfogalactosylceramide) OL-2: applications of antigen immunodetections in brain tissues and urinary samples.
An IgM monoclonal antibody, OL-2, was produced by immunizing Lou rats with crude cerebellar membrane fraction. Splenocytes from the rats were fused with a rat myeloma cell line. An antibody secreted by one hybridoma was found to bind to sulfated glycolipids, i.e. sulfatide, seminolipid, sulfolactosylceramide, lysosulfatide and evidenced no binding to neutral sphingoglycolipids such as galactosylceramide, and lactosylceramide, as shown by immunodetection by thin-layer chromatography. In tissue sections, cerebellar white matter and oligodendrocytes were strongly labeled while live; immunocytofluorescence detected both immature and fully mature oligodendrocyte in tissue cultures. The antibody was successfully used to detect urinary sulfatides in metachromatic leukodystrophy and distinguish them from closely migrating other lipids from patients with other neurological diseases.
Colsch
Benoit
B
Université Pierre et Marie Curie UMR CNRS 7613, 4 Place Jussieu, 75252 Paris cedex 05, France.
Baumann
Nicole
N
Ghandour
M Said
MS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D015153:Blotting, Western; D001923:Brain Chemistry; D002855:Chromatography, Thin Layer; D000939:Epitopes; D006801:Humans; D007150:Immunohistochemistry; D007966:Leukodystrophy, Metachromatic / Q000652:urine*; D051381:Rats; D013433:Sulfoglycosphingolipids / Q000032:analysis / Q000276:immunology* / Q000652:urine
D000911:Antibodies, Monoclonal; D000939:Epitopes; D013433:Sulfoglycosphingolipids