title: Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides.
abstract: Mice genetically engineered to lack complex gangliosides are improved hosts for raising antibodies against those gangliosides. We report the generation and characterization of nine immunoglobulin G (IgG)-class monoclonal antibodies (mAbs) raised against the four major brain gangliosides in mammals. These include (designated as ganglioside specificity-IgG subclass) two anti-GM1 mAbs (GM1-1, GM1-2b), three anti-GD1a mAbs (GD1a-1, GD1a-2a, GD1a-2b), one anti-GD1b mAb (GD1b-1), and three anti-GT1b mAbs (GT1b-1, GT1b-2a, GT1b-2b). Each mAb demonstrated high specificity, with little or no cross-reactivity with other major brain gangliosides. Enzyme-linked immunosorbent assay (ELISA) screening against 14 closely related synthetic and purified gangliosides confirmed the high specificity, with no significant cross-reactivity except that of the anti-GD1a mAbs for the closely related minor ganglioside GT1a alpha. All of the mAbs were useful for ELISA, TLC immunooverlay, and immunocytochemistry. Neural cells from wild-type rats and mice were immunostained to differing levels with the anti-ganglioside antibodies, whereas neural cells from mice engineered to lack complex gangliosides (lacking the ganglioside-specific biosynthetic enzyme UDP-GalNAc:GM3/GD3 N-acetylgalactosaminyltransferase) remained unstained, demonstrating that most of the mAbs react only with gangliosides and not with related structures on glycoproteins. These mAbs may provide useful tools for delineation of the expression and function of the major brain gangliosides and for probing the pathology of anti-ganglioside autoimmune diseases.
authors.affiliation: Department of Pharmacology and Molecular Sciences, The Johns Hopkins School of Medicine, 725 N. Wolfe Street, Baltimore, Maryland 21205-2185, USA. schnaar@jhu.edu
authors.forename: Ronald L
authors.initials: RL
authors.lastname: Schnaar
authors.forename: Susan E
authors.initials: SE
authors.lastname: Fromholt
authors.forename: Yanping
authors.initials: Y
authors.lastname: Gong
authors.forename: Alka A
authors.initials: AA
authors.lastname: Vyas
authors.forename: Wouter
authors.initials: W
authors.lastname: Laroy
authors.forename: Dawn M
authors.initials: DM
authors.lastname: Wayman
authors.forename: Marija
authors.initials: M
authors.lastname: Heffer-Lauc
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ito
authors.forename: Hideharu
authors.initials: H
authors.lastname: Ishida
authors.forename: Makoto
authors.initials: M
authors.lastname: Kiso
authors.forename: John W
authors.initials: JW
authors.lastname: Griffin
authors.forename: Kazim A
authors.initials: KA
authors.lastname: Shiekh
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D001665:Binding Sites; D002531:Cerebellum / Q000737:chemistry*; D005679:G(M3) Ganglioside / Q000276:immunology; D005732:Gangliosides / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice
chemical_list: D000911:Antibodies, Monoclonal; D005679:G(M3) Ganglioside; D005732:Gangliosides; D007074:Immunoglobulin G; C021298:ganglioside, GD1a; C033851:ganglioside, GD1b; C016481:trisialoganglioside GT1