Dot-blot immunodetection of antibodies against GM1 and other gangliosides on PVDF-P membranes.
A simple and rapid assay for detection of antibodies against GM1 and other gangliosides (GM3, GM2, GD1a, GD1b, GT1b, GD3) is described. Purified gangliosides were applied individually in 1 microliter of methanol to polyvinylidene difluoride (PVDF) membranes. Anti-ganglioside antibodies in human sera were allowed to bind and were revealed with a second antibody coupled to peroxidase. The specificity of antibodies binding to gangliosides was confirmed using established techniques to detect anti-ganglioside antibodies such as immunostaining of gangliosides after high performance thin layer chromatography according to Derrington et al. (1989) and ELISA procedure according to Adams et al. (1991) or using the ability of cholera toxin beta subunit to remove GM1 bound antibodies. The dot-blot assay is the simplest and quickest method to run and it appears to be suitable for large routine screening detection of anti-ganglioside antibodies in sera of patients with neurological diseases.
Chabraoui
F
F
Laboratoire d'Immunobiologie, Service Pr. Quincy, Hôpital Neurologique, Lyon, France.
Derrington
E A
EA
Mallie-Didier
F
F
Confavreux
C
C
Quincy
C
C
Caudie
C
C
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000378:metabolism; D001667:Binding, Competitive; D001923:Brain Chemistry; D002772:Cholera Toxin / Q000378:metabolism / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004797:Enzyme-Linked Immunosorbent Assay; D005677:G(M1) Ganglioside / Q000276:immunology*; D005732:Gangliosides / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D046911:Macromolecular Substances; D008567:Membranes, Artificial; D011145:Polyvinyls; D051381:Rats; D012680:Sensitivity and Specificity; D005465:Fluorocarbon Polymers
D000906:Antibodies; D002772:Cholera Toxin; D005677:G(M1) Ganglioside; D005732:Gangliosides; D046911:Macromolecular Substances; D008567:Membranes, Artificial; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers