Recovery of gangliosides from aqueous solutions on styrene-divinylbenzene copolymer columns

Abstract

An investigation was made using a styrene-divinylbenzene copolymer as a solid phase sorbent to recover gangliosides from aqueous solutions. A comparison with octadecyl-bonded (C18) silica gel showed that the general procedure used to purify gangliosides on C18 silica gel could be used with the copolymer. The yield of gangliosides depended on various parameters such as the composition of the conditioning solution, the salt concentration of the loading solution, and the amount of applied gangliosides per gram of copolymer. In optimal conditions, the recovery of gangliosides and other lipids present in the upper phases of partition was higher than 95%. Using radiolabeled gangliosides, it was found that gangliosides present in serum-containing medium could also be quantitatively recovered on copolymer, provided the medium was diluted with an equal volume of methanol prior to its application onto the column.The major advantage of the copolymer is its high stability in acidic or alkaline conditions that allows multiple cycles of cleaning and reconditioning of the sorbent without alteration of its chromatographic properties.

The mitochondria-associated endoplasmic-reticulum subcompartment (MAM fraction) of rat liver contains highly active sphingolipid-specific glycosyltransferases

“…The reaction products of the SAT assays were extracted with chloroform/methanol (1:1, v/v), filtered, evaporated to dryness and the gangliosides were freed of contaminants as described by Popa et al [28]. The dry residue was taken up in 2 ml of methanol/PBS, pH 7.4 (1/1, v/v) and the material applied to a 3 ml styrene/divinylbenzene column equilibrated in the same solvent.…”

“…Again, the assay was performed using Lac-Cer adsorbed on to silica gel as an exogenous acceptor incubated for 2 h in the presence of CMP-[ 14 C]N-acetylneuraminate. After purification of the gangliosides with our recently published new procedure [28], the gangliosides were resolved by TLC using solvent III. Figure 5 shows the results obtained in a representative experiment.…”

Chimeric Antibody c.8B6 to O-Acetyl-GD2 Mediates the Same Efficient Anti-Neuroblastoma Effects as Therapeutic ch14.18 Antibody to GD2 without Antibody Induced Allodynia

“…The gangliosides were then obtained from the pooled upper phase from the Folch partitions by reverse-phase chromatography on styrene divinylbenzene copolymer columns (EnviChrom™, Supelco, L’isle D’Abeau, France) as described by Popa et al. [19] . In some experiments, the O-acetyl groups from gangliosides were removed by alkaline treatment with concentrated ammonium hydroxide overnight at 25°C.…”

Recent advances in expanding the coverage of the lipidome

“…More recently, extraction procedures with less hazardous solvents have been introduced [ 17 ]. Finally, several protocols for the extraction of more hydrophilic lipid species have been published [ 25 , 26 , 27 ].…”