Identity of calcitonin extracted from normal human thyroid glands with synthetic human calcitonin-(1-32).
Calcitonin in human thyroid glands obtained at autopsy from normal subjects were extracted with 2 M acetic acid. The extracts were additionally purified by adsorption to Sep-Pak C18 cartridges, and calcitonin was identified after gel filtration analysis, reverse-phase high-performance liquid chromatography (HPLC), thin-layer chromatography and isoelectric focusing. The purification steps were monitored by radioimmunoassay, and partially purified calcitonin was used for biological and physicochemical comparison with synthetic human calcitonin-(1-32) and its Met8-sulfoxide form. On gel filtration analysis a predominant peak coeluted with the synthetic hormone, and on HPLC two discrete peaks with the retention times of monomeric and dimeric human calcitonin were found. Thin-layer chromatography allowed the detection of two peaks with the Rf of human calcitonin-(1-32) and of its sulfoxide, respectively. The pI (7.9) of the predominant peaks of synthetic calcitonin were identical. Our findings provide strong evidence that the predominant forms of human calcitonin extracted from normal thyroid glands correspond to synthetic calcitonin-(1-32) and to dimeric calcitonin.
D002116:Calcitonin / Q000138:chemical synthesis / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D006801:Humans; D011863:Radioimmunoassay; D013961:Thyroid Gland / Q000032:analysis*
D002116:Calcitonin