Rajkowski K M, Cittanova N, Desfosses B, Jayle M F
Steroids. 1977 May;29(5):701-13. doi: 10.1016/0039-128x(77)90021-6.
The formation of a horseradish peroxidase-testosterone conjugate for the enzyme-linked immunoassay of testosterone was investigated, using tritiated testosterone to follow the reaction. The formation of testosterone-3-(carboxymethyl) oxime-peroxidase by the mixed anhydride method was found to give a conjugate of high enzymatic activity and with three molecules of testosterone per molecule of peroxidase. The optimum conditions for the assay of peroxidase activity were studied and an assay capable of measuring 1 to 5 ng of the conjugate developed; the standard curve being virtually linear. The stability of the conjugate in solution and the effect of lyophilisation on enzymatic activity are also described. The peroxidase-testosterone conjugate was suitable for enzyme-linked immunoassay and the quantities measurable with the peroxidase assay covered the range necessary for a plasma testosterone assay. The stability of the conjugate was such that no particular precautions were necessary for its storage.
研究了用于睾酮酶联免疫分析的辣根过氧化物酶-睾酮偶联物的形成，使用氚标记的睾酮追踪反应。发现通过混合酸酐法形成睾酮-3-(羧甲基)肟-过氧化物酶可得到具有高酶活性且每分子过氧化物酶含有三个分子睾酮的偶联物。研究了过氧化物酶活性测定的最佳条件，并开发了一种能够测量1至5 ng偶联物的测定方法；标准曲线几乎呈线性。还描述了偶联物在溶液中的稳定性以及冻干对酶活性的影响。过氧化物酶-睾酮偶联物适用于酶联免疫分析，过氧化物酶测定法可测量的量涵盖了血浆睾酮测定所需的范围。偶联物的稳定性使得其储存无需特别的预防措施。
