Radioimmunoassay of cholecystokinin-pancreozymin

Abstract

A sensitive and specific radioimmunoassay for cholecystokinin-pancreozymin (CCK-PZ) has been developed, using rabbit antisera to crude porcine hormone. Highly purified porcine CCK-PZ, labelled with 131I, and repurified by column chromatography on Sephadex G15, was used as tracer. Separation of free from antibody-bound labelled CCK-PZ was carried out using charcoal, ion-exchange resin, or a double antibody procedure. Non-specific interference with the assay system by serum factors was abolished (as judged by in-vitro and in-vivo recovery studies) by boiling and diluting the serum samples before assay. Ninety-nine per cent pure porcine CCK-PZ (standard), commercial CCK-PZ preparations, caerulein, the C-terminal 8- and 12-amino acid fragments of the CCK-PZ molecule, and endogenous human CCK-PZ all cross reacted in the assay system and showed parallel inhibition curves. No significant cross reaction was found with gastrin, secretin, glucagon, or insulin. The sensitivity of the assay is approximately 5 pg per ml of test solution, which proved adequate for measuring physiological levels of CCK-PZ in peripheral blood in man. The mean immunoreactive CCK-PZ concentration in 50 fasting normal subjects was 60·4 pg per ml. The distribution of individual values was skewed, however, so that the median was much lower (30 pg per ml). Older subjects had higher fasting levels of CCK-PZ than were found in young adults.