Autoantigen microarrays for multiplex characterization of autoantibody responses.
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.
Robinson
William H
WH
Division of Immunology and Rheumatology, Department of Medicine, Stanford, California, USA. wrobins@stanford.edu
DiGennaro
Carla
C
Hueber
Wolfgang
W
Haab
Brian B
BB
Kamachi
Makoto
M
Dean
Erik J
EJ
Fournel
Sylvie
S
Fong
Derek
D
Genovese
Mark C
MC
de Vegvar
Henry E Neuman
HE
Skriner
Karl
K
Hirschberg
David L
DL
Morris
Robert I
RI
Muller
Sylviane
S
Pruijn
Ger J
GJ
van Venrooij
Walther J
WJ
Smolen
Josef S
JS
Brown
Patrick O
PO
Steinman
Lawrence
L
Utz
Paul J
PJ
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
D001323:Autoantibodies; D001324:Autoantigens; D005456:Fluorescent Dyes; D007132:Immunoglobulin Isotypes