Biological potency and radioimmunoassay of canine calcitonin

H. A. W. Hazewinkel, I. Schoenmakers, D. Pelling, M. Snijdelaar, J. Wolfswinkel, J. A. Mol

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Abstract

Calcitonin (CT) is a major calcitropic hormone. Because of low cross reactivity of canine CT (cCT) in radioimmunoassays (RIA) developed for other species, a homologous RIA is needed. Synthesis of cCT allowed study of its biologic potency using a rat bioassay and its plasma half-life in dogs. The availability of cCT also made possible the development of a homologous RIA for measurement of basal and stimulated plasma CT concentrations in dogs. The biologic potency of the synthesized cCT in rats is 24 IU/mg of peptide, which is low in comparison with the 4,000 IU/mg of the salmon CT standard. In the dog, an even lower potency of 4.4 IU/mg of cCT was found. Measurement of the disappearance of iv-injected radioiodinated or nonradioiodinated cCT revealed a short biologic half-life of less than 3 min, followed by a long half-life of 20 min. A polyclonal antiserum against synthetic cCT was raised in a goat. Using a final antiserum dilution of 1:12,000 and 125I-labeled synthetic cCT, the RIA had a detection limit of 6.5 ng/l. The antibody did not crossreact with standard human CT and had <0.1% cross reactivity with porcine CT. For measurement of plasma cCT concentrations, an extraction procedure was developed using ethanol. Dilutions of synthetic cCT and canine plasma extracts revealed parallelism over a wide range of concentrations. Size exclusion chromatography of canine plasma extracts on Biogel P-10 revealed a single cCT peak at the same position as [125I]-cCT, showing that there was little interference by other proteins or cCT prohormone. Basal plasma CT concentrations were 12–80 ng/l, and there was an 8- and 20-fold increase after calcium (1 and 2.5 mg/kg body weight) bolus infusion.
Original languageEnglish
Pages (from-to)333-344
Number of pages12
JournalDomestic Animal Endocrinology
Volume17
Issue number4
DOIs
Publication statusPublished - 1 Nov 1999

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