[The research of radioimmunoassay using double Abs for bradykinin]

Bradykinin (BK) is a potent vasodilative substance, and plays great physiological and pathological roles in animals and human beings. To measure the quantity of BK, the radioimmunoassay (RIA) has been devised, but the traditional RIA method has certain defects, such as presence of numerows interfering factors and errors and time consuming. Now, we produce anti-BK serum in rabbits by using BK-ovalbumin conjugate as an immunogen, and the 125I labeled Tyr8-BK by using a modified chloramine-T method. High specific activity has been obtained after purification with DEAE-Sephadex A-25 column chromatography. We use the donkey anti-rabbit Ab and PEG 6000 to separate the bound from the free 125I-Tyr8-BK. The limitation range of standard curve is from 25 to 1600 pg, NSB is 3.1% affinity constant (K) is 0.8 x 10(10) L/mol, and there is no significant interference with other biological BK analogues. The blood samples are treated by adding Polybrene (inhibitor) and PEG 6000 to deposit the big serum proteins in order to reduce the disturbing substances. This method has been shown to be a sensitive, specific, reliable, simple and convenient measure of the serum BK level. By this method, the serum BK quantities in men, women and rats are respectively 1584 +/- 347 pg/ml, 1642 +/- 302 pg/ml and 1805 +/- 225 pg/ml, the recycling rate is 95%, the intergroup CV is 5.0% and outergroup CV = 9.2%.