Gel chromatography of immunoextracted plasma calcitonin in response to the calcium clamp in healthy males

Abstract

Abstract. A radioimmunoassay (RIA) was developed for immunoreactive calcitonin (iCT) in human plasma. Antibodies against synthetic human calcitonin (hCT) coupled to bovine serum albumin (BSA) were raised in rabbits and were directed against the carboxy terminal part of CT. The detection limit of the assay was 8 pg/ml. In 7 males the iCT response to a calcium-clamp was studied. Blood was collected at 0, 30 and 60 min after the start of the calcium infusion. iCT was measured directly in plasma and in extracts obtained after purification of plasma iCT by means of immobilized CT antibodies. There was a good correlation between iCT in plasma samples and extracts, r = 0.993, n = 14 (P < 0.001). Dilution curves of extracts and plasma were parallel with the hCT standard curves. Gel chromatography of the extracts on Sephadex G-50 and G-75 disclosed heterogeneity of iCT in normal plasma during basal conditions as well as during calcium stimulation. Thirty min after the start of the calcium clamp all molecular forms, most likely constituting monomeric and dimeric CT and larger forms, were increased, while after 60 min iCT seemed to constitute predominantly forms larger than monomeric CT. Basal levels of unextracted iCT in healthy males (n = 44, 37 ± 10 years) were 15 ± 9 pg-equivalents/ml (mean ± sd) which was higher than in females (n = 40, 32 ± 9 years) 11 ± 4 pg-equivalents/ml (P < 0.05).