Radioimmunoassay of human plasma somatostatin.

Abstract

Human plasma somatostatin-like immunoreactivity (SLI) is heterogeneous. Moreover, human plasma contains a somatostatin-binding protein, enzymes that degrade the peptide, and other poorly characterized substances that interfere in radioimmunoassays of the peptide. Previously described extraction methods only partially circumvent these problems and generally result in less than adequate recovery. The present studies were therefore undertaken to determine whether gel filtration of plasma could be used to avoid these difficulties and provide a means for radioimmunoassay of plasma somatostatin in man. Gel filtration (Sephadex G25) of human plasma and subsequent radioimmunoassay of column eluates using five different antisera uniformly demonstrated two fractions of SLI in plasma. One fraction (peak I) eluted in the void volume and had an apparent molecular weight in excess of 150,000 daltons; this material comprised most of the plasma SLI, did not dilute in parallel with synthetic somatostatin, and decreased immunoprecipitability of both N-tyrosine- and tyrosine-11-125I somatostatin. The second fraction (peak II) coeluted with synthetic somatostatin (apparent mol. wt. 1600 daltons), diluted in parallel with synthetic somatostatin, and did not appreciably degrade either 125I-tyrosylated analogue. Synthetic somatostatin added to plasma was recovered nearly completely (95 ± 3%) in peak II SLI. Infused synthetic somatostatin increased peak II SLI but did not alter peak I SLI. These results demonstrate that gel chromatography of human plasma before assay can be used to circumvent problems of immunoheterogeneity, 0degradation, and nonspecific interference while providing adequate recovery. With this method, mean plasma somatostatin concentrations (peak II SLI) in postabsorptive normal volunteers ranged from approximately 40 to 100 pg/ml, depending on the antibody system used. These concentrations (3–6 × 10−11 M) are of similar magnitude to thosereported to affect reputed target tissues of somatostatin in vitro and are thus consistent with a possible hormonal function for the peptide.