Abstract
Type 1 diabetes (T1D) is an autoimmune disease which results from the destruction of pancreatic beta cells. Autoantibodies directed against islet antigens are valuable diagnostic tools. Insulin autoantibodies (IAAs) are usually the first to appear and also the most difficult to detect amongst the four major islet autoantibodies. A non-radioactive IAA bridging ELISA was developed to this end. In this assay, one site of the IAAs from serum samples is bound to a hapten-labeled insulin (GC300-insulin), which is subsequently captured on anti-GC300 antibody-coated 96-well plates. The other site of the IAAs is bound to biotinylated insulin, allowing the complex to be detected by an enzyme-streptavidin conjugate. In the present study, 50 serum samples from patients with newly diagnosed T1D and 100 control sera from non-diabetic individuals were analyzed with our new assay and the results were correlated with an IAA radioimmunoassay (RIA). Using IAA bridging ELISA, IAAs were detected in 32 out of 50 T1D children, whereas with IAA RIA, 41 out of 50 children with newly diagnosed T1D were scored as positive. In conclusion, the IAA bridging ELISA could serve as an attractive approach for rapid and automated detection of IAAs in T1D patients for diagnostic purposes.
Highlights
Type 1 diabetes (T1D) is an autoimmune disease characterized by the destruction of insulin-producing pancreatic beta cells within the islets of Langerhans
Insulin autoantibodies (IAAs) are usually the first to appear before T1D development and they are most frequently found in young children, as their level and prevalence at diagnosis inversely correlate with age [2]
We describe the development of a non-radioactive IAA assay using a bridging enzyme-linked immunosorbent assay (ELISA) format, which could be a valid alternative to RIAs routinely used in most clinical laboratories
Summary
Type 1 diabetes (T1D) is an autoimmune disease characterized by the destruction of insulin-producing pancreatic beta cells within the islets of Langerhans During this autoimmune process, autoantibodies are generated that react against several beta-cell antigens, e.g. insulin, glutamic acid decarboxylase (GAD65), protein tyrosine phosphatase (IA-2) and zinc transporter 8 (ZnT8). Autoantibodies are generated that react against several beta-cell antigens, e.g. insulin, glutamic acid decarboxylase (GAD65), protein tyrosine phosphatase (IA-2) and zinc transporter 8 (ZnT8) These autoantibodies can be present years before disease onset [1], allowing for an early diagnosis before clinical manifestations. Recent studies have used electrochemiluminescence (ECL) detection developed by Meso Scale Discovery (MSD) as a method for measuring IAAs [5,6] This technique does not require synthesis of radiolabeled antigens, dedicated equipment is needed, with a relatively high cost compared with most other technologies. There is a compelling need for new and better methods to measure IAAs in terms of sensitivity, cost and time requirements
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