Zaidi M, Chambers T J, Moonga B S, Oldoni T, Passarella E, Soncini R, MacIntyre I
Department of Chemical Pathology, Royal Postgraduate Medical School, London, U.K.
J Endocrinol Invest. 1990 Feb;13(2):119-26. doi: 10.1007/BF03349520.
We report the development and validation of three microbioassays for calcitonin based on calcitonin-induced inhibition of the activity of isolated osteoclasts. Having precisely quantified osteoclast motility, spreading and bone resorptive activity, we have applied stringent analytical procedures to define assay characteristics. We have found that the appropriately transformed responses significantly regress on log dose of the peptides. Furthermore, potency estimates obtained using calcitonins from three species (human, salmon and a synthetic analogue of eel calcitonin) have been found to be consistent with those obtained using conventional calcitonin bioassays. In addition, the assays are remarkably sensitive (detection limit 10(-15) M), highly specific and precise. We have determined plasma levels of bioactive calcitonin on samples from patients with medullary thyroid carcinoma; these are several-fold lower than those obtained using our routine calcitonin radioimmunoassay. Our study thus, forms the basis of an entirely new approach for the determination of 'biologically active' calcitonin, and we envisage that such target cell-specific assays could become useful microanalytical methods.
我们报告了基于降钙素诱导分离破骨细胞活性抑制的三种降钙素微生物测定法的开发与验证。在精确量化破骨细胞的运动性、铺展性和骨吸收活性后，我们应用了严格的分析程序来确定测定法的特性。我们发现，经过适当转换的反应与肽的对数剂量显著回归。此外，使用来自三种物种（人类、鲑鱼和鳗鱼降钙素的合成类似物）的降钙素获得的效价估计值与使用传统降钙素生物测定法获得的估计值一致。此外，这些测定法非常灵敏（检测限为10^(-15) M）、高度特异且精确。我们测定了甲状腺髓样癌患者样本中生物活性降钙素的血浆水平；这些水平比使用我们常规的降钙素放射免疫测定法获得的水平低几倍。因此，我们的研究构成了一种全新的测定“生物活性”降钙素方法的基础，并且我们设想这种靶细胞特异性测定法可能会成为有用的微量分析方法。
