DeWailly D, Maes M, Amrhein J A, Rothwell S W, Migeon C J
J Lab Clin Med. 1979 Jul;94(1):106-13.
We have adapted the method of Castañeda and Liao to the assay of DHT receptors in cultured fibroblasts arising from human skin. An antitestosterone antibody (100% crossreactivity for DHT) was coupled to CNBr-activated Sepharose. Confluent monolayers of fibroblasts were incubated with 3H-DHT (2 nM) at 37 degrees C for 30 min. Fibroblasts were then collected, sonicated, and centrifuged at 1200 x g for 15 min. The receptor assay was carried out on the supernatant; the antibody-sepharose was used to remove both unbound and nonspecifically bound DHT. Experience showed that the antibody did not entirely remove the nonspecifically bound and free DHT. A blank (sample heated at 60 degrees C for 3 min) was therefore subtracted to obtain an accurate value of specifically bound DHT. In spite of this, the antibody method, when compared to the gel filtration method, was more rapid and more convenient. Its reproducibility was similar to that of the gel filtration method, and its sensitivity was somewhat greater in patients with low levels of DHT-receptor complex. Improved sensitivity could be particularly useful when dealing with partial AIS.
我们已将卡斯塔涅达和廖的方法应用于检测源自人皮肤的培养成纤维细胞中的双氢睾酮（DHT）受体。将一种抗睾酮抗体（对DHT的交叉反应性为100%）偶联到溴化氰活化的琼脂糖上。将成纤维细胞汇合单层在37℃下与3H-DHT（2 nM）孵育30分钟。然后收集成纤维细胞，超声处理，并在1200×g下离心15分钟。在上清液中进行受体检测；抗体-琼脂糖用于去除未结合的和非特异性结合的DHT。经验表明，该抗体不能完全去除非特异性结合的和游离的DHT。因此，减去一个空白（样品在60℃加热3分钟）以获得特异性结合的DHT的准确值。尽管如此，与凝胶过滤法相比，抗体法更快且更方便。其重现性与凝胶过滤法相似，并且在DHT受体复合物水平较低的患者中其灵敏度略高。在处理部分雄激素不敏感综合征（AIS）时，提高的灵敏度可能特别有用。
