A microarray enzyme-linked immunosorbent assay for autoimmune diagnostics

• Published in: Electrophoresis Vol. 21; no. 13; pp. 2641 - 2650
• Main Authors: Joos, Thomas O., Schrenk, Monika, Höpfl, Peter, Kröger, Kerstin, Chowdhury, Ushashi, Stoll, Dieter, Schörner, Dominik, Dürr, Manfred, Herick, Klaus, Rupp, Steffen, Sohn, Kai, Hämmerle, Hugo
• Format: Journal Article
• Language: English
• Published: Hoboken Wiley Subscription Services, Inc., A Wiley Company 01.07.2000
• Subjects: Antibodies, Antinuclear - blood; Autoantibodies - blood; Autoantigens - immunology; Autoimmune disease; Autoimmune Diseases - blood; Autoimmune Diseases - diagnosis; Autoimmune Diseases - immunology; Biotinylation; Chemiluminescence; Dose-Response Relationship, Immunologic; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - instrumentation; Enzyme-Linked Immunosorbent Assay - methods; Humans; Immunoassay; Immunoglobulin G - immunology; Luminescent Measurements; Microarray; Microchemistry - instrumentation; Microchemistry - methods; Recombinant Fusion Proteins - immunology; Replica Techniques; Sensitivity and Specificity
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/1522-2683(20000701)21:13<2641::AID-ELPS2641>3.0.CO;2-5

In order to quantify autoantibodies in the sera of patients with autoimmune disease, we have created a microarray‐based immunoassay that allows the simultaneous analysis of 18 known autoantigens. The microarrays contain serial dilutions of the various antigens, thereby allowing accurate determination of autoantibody titer using minimal amounts of serum. The assay is very sensitive and highly specific: as little as 40 fg of a known protein standard can be detected with little or no cross‐reactivity to nonspecific proteins. The signal intensities observed from serial dilutions of immobilized antigen correlate well with serial dilutions of autoimmune sera. Miniaturized and highly parallelized immunoassays like these will reduce costs by decreasing reagent consumption and improve efficiency by greatly increasing the number of assays that can be performed with a single serum sample. This system will significantly facilitate and accelerate the diagnostics of autoimmune diseases and can be adapted easily to any other kind of immunoassay.