title: Immunological characterization of antibodies against synthetic peptides of glutamic acid decarboxylase.
abstract: We characterized antibodies against synthetic N-terminal peptides (glutamic acid decarboxylase; GAD65N and GAD67N) and a C-terminal peptide (GAD67C) of human GAD isoforms. On Western blots, the GAD65N antibody specifically stained the 65 kDa isoform and the GAD67N antibody the 67 kDa one in various mammalian brain tissues, whereas the GAD67C antibody stained both. The immunotrapped GAD enzyme activity increased in a dose-dependent manner with increasing concentration of the N-terminal peptide antibodies, but the activity was completely inhibited by the C-terminal peptide antibody. By an enzyme-linked immunosorbent assay using rat brain GAD purified on a GAD67C antibody-affinity column, we detected GAD antibodies in 40% (24/60) of the patients with long-standing insulin-dependent diabetes mellitus (IDDM). These antipeptide antibodies are a useful tool not only for identifying the GAD isoforms, but also for purifying GAD.
authors.affiliation: Department of Biochemistry, Utano National Hospital, Kyoto, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Ohta
authors.forename: H
authors.initials: H
authors.lastname: Obayashi
authors.forename: T
authors.initials: T
authors.lastname: Ichimura
authors.forename: M
authors.initials: M
authors.lastname: Nishimura
authors.forename: N
authors.initials: N
authors.lastname: Itoh
authors.forename: K
authors.initials: K
authors.lastname: Ohta
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002417:Cattle; D003922:Diabetes Mellitus, Type 1 / Q000201:enzymology; D005968:Glutamate Decarboxylase / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D051379:Mice; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D005968:Glutamate Decarboxylase