Brown W B, Peacock M
Clin Chim Acta. 1986 Sep 15;159(2):111-21. doi: 10.1016/0009-8981(86)90043-4.
Antisera to 1,25-dihydroxycholecalciferol were raised to 1,25-dihydroxycholecalciferol 25-hemisuccinate-bovine serum albumin, 1,25-dihydroxycholecalciferol 3-hemisuccinate-bovine serum albumin, 1,25-dihydroxycholecalciferol 3-hemisuccinate-porcine thyroglobulin and (5Z,7E)-(1S,3R)-1,3 dihydroxy-9,10-seco-24,25, 26,27-tetrakisnor-5,7,10(19)-cholestatrien-23-oic acid porcine thyroglobulin in rabbits. The antisera cross-reacted with a wide spectrum of vitamin D metabolites but their affinity was highest for 1,25-dihydroxycholecalciferol and some differentiated between ergo- and cholecalciferol metabolites. The presence of vitamin D binding protein and the pH of incubation markedly affected the sensitivity and specificity of antisera. A number of antisera were capable of measuring the normal plasma concentrations of vitamin D metabolites and their different affinities could be used for the measurement of 1,25-dihydroxyergocalciferol and 1,25-dihydroxycholecalciferol in plasma. None however were specific enough for the direct measurement of the metabolites in plasma. Immunisation increased the plasma concentration of endogenous 1,25-dihydroxyergo- and cholecalciferol in the animals.
针对1,25 - 二羟胆钙化醇制备了抗血清，所用抗原分别为1,25 - 二羟胆钙化醇25 - 半琥珀酸酯 - 牛血清白蛋白、1,25 - 二羟胆钙化醇3 - 半琥珀酸酯 - 牛血清白蛋白、1,25 - 二羟胆钙化醇3 - 半琥珀酸酯 - 猪甲状腺球蛋白以及(5Z,7E)-(1S,3R)-1,3 - 二羟基 - 9,10 - 开环 - 24,25,26,27 - 四去甲 - 5,7,10(19)-胆甾三烯 - 23 - 酸猪甲状腺球蛋白，免疫动物为兔。这些抗血清与多种维生素D代谢产物发生交叉反应，但其对1,25 - 二羟胆钙化醇的亲和力最高，且能区分麦角钙化醇和胆钙化醇的代谢产物。维生素D结合蛋白的存在以及孵育时的pH值显著影响抗血清的敏感性和特异性。一些抗血清能够检测维生素D代谢产物的正常血浆浓度，其不同的亲和力可用于检测血浆中的1,25 - 二羟麦角钙化醇和1,25 - 二羟胆钙化醇。然而，没有一种抗血清特异性足以直接检测血浆中的代谢产物。免疫接种使动物体内内源性1,25 - 二羟麦角钙化醇和1,25 - 二羟胆钙化醇的血浆浓度升高。
