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Properties of Antisera to Ganglioside G_M1 and Asialo G_M1^1,2,

From the Departments of Medicine, Microbiology and Immunology, and Neurology, Albert Einstein College of Medicine, Bronx, New York 10461

Abstract

The object of this study is to prepare antisera to purified glycosphingolipids for immunocytochemical studies. Rabbits were immunized with ganglioside G_M1 complexed either to methylated BSA or to a glycoprotein isolated from human erythrocytes, and to asialo G_M1 complexed to the erythrocyte glycoprotein. IgG and IgM antibody fractions were separated by chromatography on Sephadex G-200 and their properties were determined by complement fixation. IgG antibodies to asialo G_M1 were highly specific and exhibited little cross-reactivity with G_M1 or other glycolipids, but IgM antibodies cross-reacted extensively with gangliosides G_M1 and G_D1b. Complement fixation of IgG anti-asialo G_M1 with asialo G_M1 was completely inhibited by about 15 nmoles of ganglio-N-tetraose, the carbohydrate moiety of this glycolipid.

Both IgG and IgM antibodies to G_M1 cross-reacted extensively with asialo G_M1 and G_D1b, but specific IgG antibodies to G_M1 were revealed by absorption with asialo G_M1. The specific and cross-reacting IgG antibodies to G_M1 were inhibited by the pentaose moiety of this glycoplipid, but these antibodies were much harder to inhibit than IgG anti-asialo G_M1. These results demonstrate that it is possible to prepare specific antisera to purified glycolipids with a single administration of antigen, in contrast to previous studies in which repeated injections of ganglioside mixtures were employed.

Footnotes

¹ Presented in part at the 57th Annual Meeting of the Federation of American Societies for Experimental Biology in Atlantic City, New Jersey, April 1973.

² This work was supported by United States Public Health Service Grants AI 05336 and NS 04834 and by Research Grant 842-A-5 from the National Multiple Sclerosis Society.

³ Career Scientist of the Health Research Council of the City of New York, I-280.