DEVELOPMENT AND EVALUATION OF A CHROMATOGRAPHIC PROCEDURE FOR PARTIAL-PURIFICATION OF SUBSTANCE-P WITH QUANTITATION BY AN ENZYME-IMMUNOASSAY

We have developed a simple chromatographic procedure for the partial p urification of substance P (SP) from acidified plasma and serum sample s. We have evaluated a sensitive antigen competition enzyme immunoassa y (EIA) for the quantitation of SP. The chromatographic procedure has recovery efficiencies ranging from 94.8 to 125%. The immunoreactivity of unknown amounts of purified SP subjected to the preparative procedu re yielded a coefficient of variance of 9.4%. The EIA yielded reproduc ible standard curves having an interassay (n = 8) correlation coeffici ent of 0.984. The evaluation of normal adult control serum yielded a m ean value of 51 pg/ml (range, 35 to 61 pg/ml). The evaluation of 3.33 x concentrates of serum-derived partially purified SP provided uncorre cted SP values of 117 to 201 pg/ml, which fell within the midpoint of the three-decalog standard curve. These studies indicate that both the preparative and quantitative procedures are required for the detectio n of SP in plasma or serum samples collected from patients with severa l clinical disorders.