Development and validation of a specific radioimmunoassay for somatostatin in human plasma

Abstract

Little is known about the factors controlling somatostatin secretion in man, and data are not available on the changes in circulating levels in various human physiological or pathophysiological states. This is mainly a consequence of the technical difficulties involved in measuring somatostatin in plasma. In the presence of plasma, binding of somatostatin tracer to antibody was consistently decreased by about 20%, and this could not be abolished by the addition of EDTA and aprotinin or by the use of specially prepared somatostatin-free plasma. Furthermore, in the presence of plasma, endogenous somatostatin does not dilute in parallel with synthetic cyclic somatostatin standard. We have, therefore, developed and validated a radioimmunoassay for somatostatin using prior extraction of the peptide onto leached silica glass. Tyrosine-II somatostatin was iodinated using lactoperoxidase and purified on ODS silica. This method is superior to iodination using chloramine-T with CMC cellulose purification, and gives a highly purified preparation with a shelf-life of at least eight weeks. Using this tracer and a specific antiserum, the limit of sensitivity of the assay was 10 pg/ml, with an intra-assay coefficient of variation of 12% (n = 16) and inter-assay coefficient of variation of 15% (n - 10). Parallelism has been demonstrated between standard synthetic cyclic somatostatin and all extracted plasma samples. The mean recovery of exogenous somatostatin from plasma was 78%. The fasting level of immunoreactive somatostatin at 0,900 hours in 40 normal subjects ranged from 17 to 81 pg/ml. Care is needed, however, when comparing these values with those obtained from other laboratories since standard preparations of somatostatin vary considerably in their immunopotency.