title: Autoantigen microarrays for multiplex characterization of autoantibody responses.
abstract: We constructed miniaturized autoantigen arrays to perform large-scale multiplex characterization of autoantibody responses directed against structurally diverse autoantigens, using submicroliter quantities of clinical samples. Autoantigen microarrays were produced by attaching hundreds of proteins, peptides and other biomolecules to the surface of derivatized glass slides using a robotic arrayer. Arrays were incubated with patient serum, and spectrally resolvable fluorescent labels were used to detect autoantibody binding to specific autoantigens on the array. We describe and characterize arrays containing the major autoantigens in eight distinct human autoimmune diseases, including systemic lupus erythematosus and rheumatoid arthritis. This represents the first report of application of such technology to multiple human disease sera, and will enable validated detection of antibodies recognizing autoantigens including proteins, peptides, enzyme complexes, ribonucleoprotein complexes, DNA and post-translationally modified antigens. Autoantigen microarrays represent a powerful tool to study the specificity and pathogenesis of autoantibody responses, and to identify and define relevant autoantigens in human autoimmune diseases.
authors.affiliation: Division of Immunology and Rheumatology, Department of Medicine, Stanford, California, USA. wrobins@stanford.edu
authors.forename: William H
authors.initials: WH
authors.lastname: Robinson
authors.forename: Carla
authors.initials: C
authors.lastname: DiGennaro
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hueber
authors.forename: Brian B
authors.initials: BB
authors.lastname: Haab
authors.forename: Makoto
authors.initials: M
authors.lastname: Kamachi
authors.forename: Erik J
authors.initials: EJ
authors.lastname: Dean
authors.forename: Sylvie
authors.initials: S
authors.lastname: Fournel
authors.forename: Derek
authors.initials: D
authors.lastname: Fong
authors.forename: Mark C
authors.initials: MC
authors.lastname: Genovese
authors.forename: Henry E Neuman
authors.initials: HE
authors.lastname: de Vegvar
authors.forename: Karl
authors.initials: K
authors.lastname: Skriner
authors.forename: David L
authors.initials: DL
authors.lastname: Hirschberg
authors.forename: Robert I
authors.initials: RI
authors.lastname: Morris
authors.forename: Sylviane
authors.initials: S
authors.lastname: Muller
authors.forename: Ger J
authors.initials: GJ
authors.lastname: Pruijn
authors.forename: Walther J
authors.initials: WJ
authors.lastname: van Venrooij
authors.forename: Josef S
authors.initials: JS
authors.lastname: Smolen
authors.forename: Patrick O
authors.initials: PO
authors.lastname: Brown
authors.forename: Lawrence
authors.initials: L
authors.lastname: Steinman
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Utz
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D001327:Autoimmune Diseases / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007132:Immunoglobulin Isotypes / Q000737:chemistry / Q000378:metabolism; D007163:Immunosorbent Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D005456:Fluorescent Dyes; D007132:Immunoglobulin Isotypes