An improved method for determination of active and total renin concentration in human plasma using an excess of sheep substrate

Abstract

A method for measurement of renin concentration (PRC) in human plasma has been developed and validated experimentally and theoretically. Like most of the previous methods, the present method is based on a radioimmunoassay of angiotensin I generated during incubation of plasma with an excess of renin substrate. In the present study we used, as the substrate, sheep angiotensinogen partially purified from anephric sheep plasma by ammonium sulfate fractionation and pepstatin-aminohexyl-agarose chromatography. The advantage of using sheep substrate is that it has an exceptionally high affinity for human renin. The partially purified substrate, which contained no detectable renin activity, improved the sensitivity, allowing quantification of renin in low-renin plasma. Possible interfering influences of plasma proteins on the radioimmunoassay were eliminated by the introduction of a simple deproteinization step. For determination of total renin concentration (TRC), inactive renin was activated by exposing plasma to low pH or trypsin. Normal values of PRC and TRC after careful selection of assay conditions were 2.9 +/- 0.4 and 33.9 +/- 6.1 ng angiotensin I X ml-1 X h-1, respectively. As an illustrative example of usefulness of the assay, PRC determination of a patient with a ectopic renin-secreting tumor is presented.