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.
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Acknowledgements
We thank K. Chong and D. Mitchell for their outstanding assistance; J.G. Lindsay for antibodies; and A. Abeliovich, H. Garren, P. Ruiz and G. Hermans for insightful discussions. This work was primarily supported by a Howard Hughes Postdoctoral Research Fellowship for Physicians Award, NIH K08 AR02133 and Arthritis Foundation Chapter Grant to W.H.R; NIH/NINDS 5R01NS18235 and NIH U19DK61934 to L.S.; and NIH K08 AI01521, an Arthritis Foundation Investigator Award, a Baxter Foundation Career Development Award, a Stanford University Office of Technology Licensing Award, and NIH U19 DK61934 to P.J.U. P.O.B is an Associate Investigator of the Howard Hughes Medical Institute.
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Robinson, W., DiGennaro, C., Hueber, W. et al. Autoantigen microarrays for multiplex characterization of autoantibody responses. Nat Med 8, 295–301 (2002). https://doi.org/10.1038/nm0302-295
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DOI: https://doi.org/10.1038/nm0302-295
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