title: Mapping of a linear autoantigenic epitope within the human thyroid peroxidase using recombinant DNA techniques.
abstract: Autoantibodies directed against the thyroid peroxidase (TPO), the thyroid microsomal antigen, are widely used to diagnose human autoimmune thyroid disease. A cloned 3.088 kb cDNA coding for the entire mature human TPO was isolated from a cDNA library derived from a pathological thyroid gland of a Graves' disease patient and used further to generate a so-called TPO epitope cDNA library in order to map linear autoantigenic epitopes involving a recombinant molecular biology approach. The TPO epitope cDNA library consisting of randomly fragmented cDNA sequences inserted in the expression vector pGEX-2T was expressed in Escherichia coli and screened with characterized anti-TPO autoantisera from Hashimoto's disease patients. All the sera were positively tested with a purified thyroid microsomal antigen fraction (TMA/TPO). Only about 1% of examined autoantisera were able to recognize bacterial expressed recombinant TPO representing sequential antigenic determinants. A corresponding autoantigenic epitope with 61 amino acids in length was located at the C-terminus of human TPO.
authors.affiliation: Department of Molecular Biology, ELIAS Entwicklungslabor, Freiburg, F.R.G.
authors.forename: B
authors.initials: B
authors.lastname: Frorath
authors.forename: C C
authors.initials: CC
authors.lastname: Abney
authors.forename: M
authors.initials: M
authors.lastname: Scanarini
authors.forename: H
authors.initials: H
authors.lastname: Berthold
authors.forename: N
authors.initials: N
authors.lastname: Hunt
authors.forename: W
authors.initials: W
authors.lastname: Northemann
mesh_terms: D001323:Autoantibodies / Q000032:analysis; D001324:Autoantigens / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D000939:Epitopes / Q000235:genetics; D006111:Graves Disease / Q000175:diagnosis; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics* / Q000276:immunology*; D010449:Peptide Mapping; D013961:Thyroid Gland / Q000201:enzymology / Q000276:immunology; D013967:Thyroiditis, Autoimmune / Q000175:diagnosis
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D004274:DNA, Recombinant; D000939:Epitopes; C056626:thyroid microsomal antibodies; D004247:DNA; D007453:Iodide Peroxidase