A sensitive enzyme immunoassay for angiotensin II in serum

A sensitive and specific enzyme immunoassay for measuring angiotensin II (AII) has been developed as a convenient alternative to a radioimmunoassay. An antiserum to AII was prepared using AII conjugated by carbodi-imide to rabbit serum albumin, and coated on to microwell plates. The labelled antigen was prepared from AII and horseradish peroxidase using the periodate method. This enzyme immunoassay was a simple two-step procedure: 0.1 ml of AII-extracted plasma was incubated for 1 h at 37 degrees C; and 1 ml of labeled AII was incubated for 1 h at 37 degrees C. Bound horseradish peroxidase activity was then determined using o-phenylenediamine as chromogen by measuring the absorbance at 492 nm. The lower detection limit of the assay was 3.5 pmol l-1. Between- and within-assay RSD values were 8.8-18.3% and 6.9-17%, respectively, for concentrations of 10-40 pmol l-1. The accuracy of the assay, determined by recovery and linearity experiments, was 89-106% for recovery and 91-126% for parallelism. The results obtained by the present ELISA method were well correlated with those obtained by an established radioimmunoassay (n = 10, r = 0.96, intercept = 0.9 and slope = 1.02). This assay is easy to perform, rapid and does not require radioisotopes; thus it could be widely applied in clinical laboratories.