Research Project
3495659
The aim of this project is to demonstrate feasibility for an immunoassay for insulin, using acridinium esters as a chemiluminescent label. This will lead to a technique for the detection of polypeptide hormones that is faster, more sensitive, and removes the need for 125I-labeled peptides or antibodies. The project will explore both traditional immunoassays where the insulin molecule is labeled, and "sandwich" type assays where insulin antibodies are used. In the first case labeled insulin will be purified by HPLC and directly substituted as the tracer in a RIA procedure. For the immunometric assay, antibodies to insulin will be labeled and purified by gel filtration chromatography. The labeled peptide and antibody will be compared for stability against a commercially available RIA using 125I-labeled insulin. The assay will serve as a model for other peptide hormone assays, which require high sensitivity and stable reagents. The project will involve limited assay development; comparison with a commercial RIA, and a limited trial with plasma samples. The multi-million dollar hormone RIA market may increase in size if more stable, safer, cheaper and simpler assays were available using chemicluminescence detection techniques.
