title: A new ELISA for dermatomyositis autoantibodies: rapid introduction of autoantigen cDNA to recombinant assays for autoantibody measurement.
abstract: Advances in immunology, biochemistry, and molecular biology have enabled the development of a number of assays for measuring autoantibodies. ELISA has been widely used, because it can deal with relatively large numbers of serum samples more quickly than other immunologic methods, such as immunoblotting and immunoprecipitation. Recombinant autoantigens, which are generally produced in E. coli using the relevant cloned cDNA, are necessary for ELISA. Conventional clinical ELISA tests are limited in their ability to purify proteins free of bacterial contaminants, and the process is labor intensive. We recently developed new ELISA tests that utilize simple in vitro transcription and translation labeling of autoantigens in order to measure dermatomyositis-(DM-) specific autoantibodies, including autoantibodies to Mi-2, MDA5, NXP-2, TIF1-α, and TIF1-γ. This method may allow for the rapid conversion of cDNAs to a chemiluminescent ELISA to detect autoantibodies that are found not only in DM but also in other autoimmune diseases.
authors.affiliation: Division of Connective Tissue Disease and Autoimmunity, Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Yoshinao
authors.initials: Y
authors.lastname: Muro
authors.affiliation: Division of Connective Tissue Disease and Autoimmunity, Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Kazumitsu
authors.initials: K
authors.lastname: Sugiura
authors.affiliation: Division of Connective Tissue Disease and Autoimmunity, Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Akiyama
mesh_terms: D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000235:genetics / Q000276:immunology; D019921:Biotinylation; D018076:DNA, Complementary; D003882:Dermatomyositis / Q000175:diagnosis* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D018076:DNA, Complementary; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins