Kobayashi N, Takama A, Shiomura K, Tabata Y, Takagi K, Shimada K
Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Steroids. 1994 Jul;59(7):404-11. doi: 10.1016/0039-128x(94)90009-4.
To obtain a group-specific antibody to 1 alpha,25-dihydroxyvitamin D and its derivatives having the 1 alpha,3 beta-dihydroxylated A-ring structure, 1 alpha-hydroxy-25,26,27-trinorvitamin D3-24-oic acid was synthesized as a haptenic derivative by a new convenient synthetic route. Three kinds of polyclonal antibodies were elicited in rabbits against the hapten conjugated with bovine serum albumin, and their specificity was examined by cross-reactivities in a radioimmunoassay procedure. One of the antibodies was found to show a suitable binding property by addition of a small amount of the plasma from vitamin D-deficient rats. Dose-response curves with measurable ranges applicable to biological specimens were obtained for 1 alpha,25-dihydroxyvitamin D3, 1 alpha,25-dihydroxyvitamin D2, (24R)-1 alpha,24,25-trihydroxyvitamin D3, (23S,25R)-1 alpha,25-dihydroxyvitamin D3 26,23-lactone, and 22-oxacalcitriol by the radioimmunoassay system using this improved antibody. The antibody will be widely useful for developing profile analyses of 1 alpha,25-dihydroxyvitamin D and its derivatives (metabolites or synthetic analogs) having the 1 alpha,3 beta-dihydroxylated A-ring, as well as immunoassays measuring each of these secosterols individually.
为获得针对1α,25 - 二羟基维生素D及其具有1α,3β - 二羟基化A环结构的衍生物的组特异性抗体，通过一种新的简便合成路线合成了1α - 羟基 - 25,26,27 - 三降维生素D3 - 24 - 油酸作为半抗原衍生物。用与牛血清白蛋白偶联的半抗原在兔体内引发了三种多克隆抗体，并通过放射免疫分析程序中的交叉反应性检测了它们的特异性。发现其中一种抗体通过添加少量维生素D缺乏大鼠的血浆表现出合适的结合特性。使用这种改进的抗体通过放射免疫分析系统获得了适用于生物标本的可测量范围内的1α,25 - 二羟基维生素D3、1α,25 - 二羟基维生素D2、(24R) - 1α,24,25 - 三羟基维生素D3、(23S,25R) - 1α,25 - 二羟基维生素D3 26,23 - 内酯和22 - 氧杂骨化三醇的剂量 - 反应曲线。该抗体将广泛用于开展对具有1α,3β - 二羟基化A环的1α,25 - 二羟基维生素D及其衍生物（代谢物或合成类似物）的谱分析，以及单独测量这些甾醇的免疫测定。
