Gangliosides and sulphatide in human cerebrospinal fluid: quantitation with immunoaffinity techniques
Davidsson, P.; Fredman, P.; Svennerholm, L.
Journal of Chromatography 496(2): 279-289
1989
ISSN/ISBN: 0021-9673 PMID: 2613833 Document Number: 336965
A senstivie micromethod involving extraction, purification and thin-layer chromatography (TLC)-enzyme immunostaining was developed for the quantation of gangliosides and sulphatide, as markers for neuronal disorders and myelin disturbances, respectively, in individual samples of less than 5 ml of cerebrospinal fluid. The gangliosides of the gangliotetraose series were individually determined with cholera toxin subunit B by TLC-enzyme-linked immunosorbent assay (ELISA) after chromatography and subsequent sialidase hydrolysis to Il3NeuAc-GgOse4Cer (GM1). Other gangliosides and sulphatide were determined with specific monoclonal antibodies by TLC-ELISA. The total ganglioside content varied between 100 and 200 nmol/l in ten normal cerebrospinal fluid samples from adults. The major gangliosides were of the gangliotetraose series, represented by GM1, IV3NeuAc,II3NeuAc-GgOse4Cer, (GD1a), Il3(NeuAc)2-GgOse4Cer (GD1b) and IV3NeuAc,Il3(NeuAc)2-GgOse4Cer (GT1b), of which the b-series gangliosides dominated, i.e., GD1b and GT1b.