Development of an enzyme-linked immunosorbent assay for serum 11-dehydrocorticosterone in rat and mouse.
Development of an enzyme-linked immunosorbent assay for serum 11-dehydrocorticosterone in rat and mouse.
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开发大鼠和小鼠血清 11-脱氢皮质酮酶联免疫吸附测定法。
DOI：
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发表时间：
2007
影响因子：
2
通讯作者：
Yoshiharu Kobayashi
中科院分区：
文献类型：
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作者：
N. Tagawa;Nao Ikariko;K. Fukumura;Yoshiharu Kobayashi
本文建立了血清11-脱氢皮质酮（4-二氢-21-醇-3，11，20-三酮）的酶联免疫吸附测定法（ELISA）。用兔抗11-脱氢皮质酮21-半琥珀酸酯标记牛血清白蛋白的抗血清。作为酶标记抗原，11-脱氢皮质酮21-半琥珀酸酯与辣根过氧化物酶结合。建立了两种ELISA系统：一种不从血清中提取类固醇（直接法），另一种使用HPLC纯化步骤（HPLC法）。除可的松（92%）外，所有测试的类固醇对抗体的交叉反应性均较低;然而，由于大鼠和小鼠中的可的松水平可忽略不计，因此可的松不会干扰该直接ELISA。直接法和HPLC法测定血清11-脱氢皮质酮的范围分别为0.3-250 ng/ml和0.78-400 ng/ml。两种方法均显示出令人满意的平行稀释、回收率和重现性;此外，每种方法获得的值与替代方法显著相关。为了评价这两种酶联免疫吸附测定系统，用这两种系统测定了正常大鼠和小鼠血清中11-脱氢皮质酮的浓度。Wistar大鼠的水平在3至14周龄之间波动（7.8+/-2.6 ng/ml），但1周龄时（1.7+/-1.2 ng/ml）明显低于其他年龄。在大鼠和小鼠中未发现性别差异。此外，使用提出的直接方法，大鼠血清11-脱氢皮质酮水平的时间变化后，11-脱氢皮质酮管理已被调查与皮质酮水平。这些结果验证了所提出的11-脱氢皮质酮的ELISA可用于测量大鼠和小鼠中11 β-HSD活性以及血清皮质酮的测定。
We have developed an enzyme-linked immunosorbent assay (ELISA) for serum 11-dehydrocorticosterone (4-pregnen-21-ol-3,11,20-trione). The antiserum against 11-dehydrocorticosterone 21-hemisuccinate-conjugated bovine serum albumin was raised in rabbits. As an enzyme-labeled antigen, 11-dehydrocorticosterone 21-hemisuccinate was conjugated to horseradish peroxidase. Two ELISA systems were established: one without the extraction of steroids from serum (direct method), and another that used an HPLC purification step (HPLC method). The cross-reactivity of all steroids tested against the antibody was low except cortisone (92%); however, since cortisone levels in rats and mice are negligible, cortisone does not interfere with this direct ELISA. The measurable range of serum 11-dehydrocortiocosterone in both the direct and HPLC methods was 0.3-250 ng/ml and 0.78-400 ng/ml, respectively. Both methods displayed satisfactory parallel dilution, recovery and reproducibility; moreover, the values obtained with each method significantly correlated with the alternate method. To evaluate the two ELISA systems, the serum concentrations of 11-dehydrocorticosterone in normal rats and mice were determined by these two systems. The levels in Wistar rats fluctuated from 3 to 14 weeks of age (7.8+/-2.6 ng/ml) but at 1 week (1.7+/-1.2 ng/ml) were significantly low compared to other ages. No sex differences were found in rats and mice. Further, using the proposed direct method, chronological changes of rat serum 11-dehydrocorticosterone levels after 11-dehydrocorticosterone administration have been investigated together with corticosterone levels. These results verify that the proposed ELISA for 11-dehydrocorticosterone is useful for measuring 11beta-HSD activities in combination with the determination of serum corticosterone in rats and mice.
人肾 11 β-羟基类固醇脱氢酶是一种高亲和力的烟酰胺腺嘌呤二核苷酸依赖性酶，与克隆的 I 型异构体不同。
DOI：
10.1210/jcem.79.2.8045966
发表时间：
1994
期刊：
The Journal of clinical endocrinology and metabolism
影响因子：
--
作者：
Stewart,PM;Murry,BA;Mason,JI
通讯作者：
Mason,JI