1. A microarray enzyme-linked immunosorbent assay for autoimmune diagnostics
- Author
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Klaus Herick, Steffen Rupp, Peter Höpfl, Hugo Hämmerle, Dominik Schörner, Ushashi Chowdhury, Kerstin Kröger, Kai Sohn, Monika Schrenk, Manfred Dürr, Dieter Stoll, Thomas O. Joos, and Publica
- Subjects
Serial dilution ,Microarray ,Recombinant Fusion Proteins ,Clinical Biochemistry ,Dose-Response Relationship, Immunologic ,Enzyme-Linked Immunosorbent Assay ,Biology ,Autoantigens ,Sensitivity and Specificity ,Biochemistry ,Autoimmune Diseases ,Analytical Chemistry ,Antigen ,medicine ,Humans ,Replica Techniques ,Biotinylation ,Autoantibodies ,Autoimmune disease ,medicine.diagnostic_test ,Microchemistry ,Autoantibody ,medicine.disease ,Molecular biology ,Titer ,Antibodies, Antinuclear ,Immunoglobulin G ,Immunoassay ,Luminescent Measurements ,DNA microarray - Abstract
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.
- Published
- 2000