title: A monoclonal antibody based enzyme-linked immunosorbent assay for the determination of GAD65, the smaller isoform of glutamic acid decarboxylase.
abstract: An enzyme-linked immunosorbent assay for GAD65, the smaller form of glutamic acid decarboxylase and an important autoantigen related to Type 1 diabetes, is described. The competitive binding assay is based on a monoclonal antibody specifically reactive with GAD65. The assay is suitable for quantification of this enzyme between 40 and 300 pg/microliter. The intraassay coefficients of variation (cv) are between 5.6% and 8.9% and the interassay cvs lie between 9.4% and 17.3%. The covalent binding of the antigen to magnetic beads as the solid phase makes the assay also applicable for quantification of GAD65 in tissue homogenates with a high concentration of detergent. The GAD65 content of islets isolated from newborn Lewis rat was detected to be 310 pg/islet. However, GAD65 was not detectable in mouse islets.
authors.affiliation: Department of Immunochemistry, Institute of Diabetes Gerhardt Katsch, Karlsburg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Hamann
authors.forename: J
authors.initials: J
authors.lastname: Hahmann
authors.forename: M
authors.initials: M
authors.lastname: Strebelow
authors.forename: B
authors.initials: B
authors.lastname: Ziegler
authors.forename: M
authors.initials: M
authors.lastname: Ziegler
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal; D001324:Autoantigens / Q000032:analysis; D001921:Brain / Q000201:enzymology*; D003922:Diabetes Mellitus, Type 1 / Q000201:enzymology / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005968:Glutamate Decarboxylase / Q000032:analysis*; D006801:Humans; D007515:Islets of Langerhans / Q000201:enzymology*; D007527:Isoenzymes / Q000032:analysis*; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008970:Molecular Weight; D051381:Rats; D011917:Rats, Inbred Lew; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D001324:Autoantigens; D007527:Isoenzymes; D005968:Glutamate Decarboxylase