Ganglioside Pattern Mapping by Gradient Overpressured Layer Chromatography

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Abstract
During the last decade overpressured layer chromatography, a forced flow planar liquid chromatographic technique has undergone significant development since its introduction in 1977. It is a high performance layer chromatographic method in its own right with varied application fields including as a substitute or a pilot technique for modern column liquid chromatography. A great advantage can be its use with gradient elution in the separation of complex biological samples shortening the laborious sample pretreatment process. The analysis of minor components is possible without overloading the chromatographic plate by the suitable selection of the polarity/solvent strength of the consecutive elution steps, overruning the major components (eluting them off the plate) before starting the final development step. Gangliosides represent a minor group of plasma membrane lipids having very important biological functions such as leukaemia and cancer antigeneity. The specific aim of the study was to perform chromatographic separation and chemical analysis of membrane gangliosides extracted from leukaemic and their normal equivalent cells, with the objective of determining and comparing the alterations in their compositions. Our step-gradient OPLC method f o r determining ganglioside patterns of several blood cell membranes, makes available a very effective sample pretreatment, the shortest sample preparation. By the results and the conclusions drawn the leukaemic and normal cells can be distinguished on the basis of GO3 expression and/or the vigorous increase in the ratio of nonpolar to polar gangliosides.