Multiplex immunoassay detection of autoimmune disease autoantibodies in serum and plasma

Abstract

Abstract Autoantibody formation is principal to the pathogenesis of a variety of autoimmune diseases. Dysregulated apoptosis and the subsequent defective clearance of cellular debris leads to the exposure of autoantigens and the generation of autoantibodies. The presence of autoantibodies may indicate disease activity, prognosis, and clinical associations related to a variety of autoimmune diseases, including systemic lupus erythematosus (SLE), Sjögren’s Syndrome, Scleroderma, Polymyositis/Dermatomyositis, and various overlap syndromes of these diseases. Here we report the development of a multiplex immunoassay to monitor 20 autoantibodies present in blood that are involved in a variety of autoimmune diseases: SSA/Ro60, SSA/Ro52, SSB/La, RNP, RNP/Sm, Sm, Ribosomal P, Proteinase 3, Myeloperoxidase, PCNA, β 2-Glycoprotein, CENP-A, CENP-B, Scl-70, Jo-1, C1q, PM/Scl-100, Ku, Mi-2, and PL-12. Using the 20-plex immunoassay, we measured expression of autoantibodies in serum and plasma from SLE patients with various disease manifestations and overlap syndromes, healthy controls, and CDC reference samples available for 11 autoantibodies. Additionally, control beads were included in the immunoassay. Levels of SSA/Ro60, SSA/Ro52, SSB/La, RNP, RNP/Sm, and Sm matched clinical laboratory features noted at the time of SLE sample collection. Additionally, anti-C1q antibody levels were associated with lupus nephritis. The CDC controls exhibited elevated MFI for the autoantibodies for which they were expected to react strongly. This study demonstrates the value of using multiplex technology to evaluate multiple autoantibodies, allowing for an extensive autoantibody profile to evaluate patients with various autoimmune diseases.