Evaluation of a commercial enzyme immunoassay for insulin in human serum, and its clinical application.

We applied a "sandwich" method, with use of beads coated with anti-insulin serum and of peroxidase-labeled anti-insulin serum, to an enzyme immunoassay of insulin in human serum. 5-Aminosalicylic acid was used as the substrate for the enzymic reaction. As little as 5 milli-int. units of insulin per liter of serum insulin was detectable. Reproducibility was satisfactory, but extraordinarily high concentrations of proinsulin and of hydrogen donors such as reduced glutathione affect results of the assay. Values determined by our enzyme immunoassay and by double-antibody radioimmunoassay correlated highly (r = 0.938, p less than 0.001, n = 216). We recommend this method for use in the clinical laboratory.