LARGE-SCALE PURIFICATION OF GANGLIOSIDES G(M3)(NEUSAC) AND G(M3)(NEU5GC) BY TRIMETHYLAMINOETHYL-FRACTOGEL HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A preparative anion-exchange high-performance liquid chromatographic m ethod for the separation of the closely allied monosialogangliosides G (M3)(Neu5Ac) and G(M3)(Neu5Gc) has been developed. Hybridoma cells, re adily available material derived from industrial monoclonal antibody p roduction, were used as ganglioside source and led to fractions with p ure G(M3)(Neu5Ac) and G(M3)(Neu5Gc) in high milligram quantities. The crude ganglioside extract was loaded onto columns filled with the stro ng anion-exchanger trimethylaminoethyl (TMAE)-Fractogel. Gangliosides were eluted from the stationary phase with a gradient system of ammoni um acetate in methanol. The scaled-up approach ranged over more than o ne order of magnitude from 20 to 500 mg batches of G(M3) gangliosides. Thus, the high-resolution power of the strong anion-exchanger TMAE-Fr actogel allowed the preparative isolation by one-step column chromatog raphy of two G(M3) specimens which only differ in one hydroxyl group a t position 5 of the neuraminic acid (N-acetyl- versus N-glycolylneuram inic acid). (C) 1998 Elsevier Science B.V. All rights reserved.