Sensitive enzyme-immunostaining and densitometric determination of ganglio-series gangliosides on thin-layer plate: pmol detection of gangliosides in cerebrospinal fluid.
An immunochemical method has been developed for sensitive detection and determination of ganglio-series gangliosides using thin-layer chromatography/enzyme-immunostaining. After chromatography of gangliosides, the plate was treated with Arthrobacter ureafaciens sialidase to remove all sialic acids from ganglio-series gangliosides. For the complete hydrolysis of gangliosides, sodium taurodeoxycholate was found to be required. The resulting asialo-glycolipids, GA2 and GA1 were reacted first with affinity-purified anti-GA2 and anti-GA1, respectively, and second with horseradish peroxidase-conjugated anti-rabbit IgG. Being highly sensitive and reproducible, it allows the characterization of gangliosides in cerebrospinal fluid which cannot be detected by classical methods.
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002855:Chromatography, Thin Layer; D005732:Gangliosides / Q000134:cerebrospinal fluid*; D006801:Humans; D007124:Immunoenzyme Techniques; D008722:Methods; D009439:Neuraminidase / Q000378:metabolism; D011817:Rabbits; D013657:Taurodeoxycholic Acid / Q000494:pharmacology
D005732:Gangliosides; D013657:Taurodeoxycholic Acid; D009439:Neuraminidase