Simplified radioimmunoassay of bradykinin in human plasma

A greatly simplified radioimmunoassay for bradykinin in human plasma is described. Current techniques require multiple chromatographic steps or extraction procedures with analytical recoveries of bradykinin of often less than 60%. We present a method in which bradykinin is separated from components of higher relative molecular mass (including kininogens) in a single step, by use of a column of Sephadex G-25 medium (PD-10). The mean analytical recovery of tritiated bradykinin added to plasma is 85.5% (SD, 3.5%). The sensitivity of this radioimmunoassay is 25 pg per assay tube, equivalent to 125 ng per liter of plasma. Twenty to 30 blood samples may be completely processed and assayed within 6 h. As determined with this technique, concentrations of bradykinin in plasma from apparently normal subjects ranged from 2.5 to 5.2 microgram/L (mean 4.2, SD 1.1 microgram/L); these values are consistent with previously reported normal values.