Aspects of bradykinin radioimmunoassay

125I-derivatives of Tyr1-kallidin, Tyr5-bradykinin, and Tyr8-bradykinin were prepared. A technique for purification of the monoiodinated derivative is described. Bradykinin antisera were tested for their ability to bind the mono-iodinated analogues. Each antiserum had a characteristic preference for one of the three labeled peptides. The sensitivity of each antiserum was greatest when it was used with the label bound most avidly by that antiserum. The specificity of an antiserum was not changed by varying the labeled analogue. Some common enzyme inhibitors had significant effects on the antigen-antibody reactions. Lecithin interfered with the reaction between antiserum and Tyr1-kallidin. The data suggest that antisera for bradykinin radioimmunoassay be tested with several radioactive iodobradykinins to maximize their usefulness. In addition, enzyme inhibitors used to stabilize levels of kinins in biological fluids should be tested for their effects on the assay. Biologic samples rich in lipid may give spurious radioimmunoassay results unless they are freed of those phospholipids that can bind labeled and unlabeled peptides.