A sensitive, robust high-throughput electrochemiluminescence assay for rat insulin
A sensitive, robust high-throughput electrochemiluminescence assay for rat insulin
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DOI：
10.1177/1087057103260515
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发表时间：
2004-02-01
影响因子：
--
通讯作者：
Seethala, R
中科院分区：
文献类型：
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作者：
Golla, R;Seethala, R
在糖尿病研究中，小鼠和大鼠模型用于体内实验，并且在不同病理生理条件下和用化合物治疗后定量血清样品中的胰岛素是必要的。很少有商业放射免疫测定和酶联免疫吸附测定（ELISA）测定试剂盒用于测定大鼠/小鼠血浆胰岛素水平。然而，使用可用的生物测定的胰岛素测量的可靠性是一个很大的问题。作者报告了一种使用Origen技术平台的稳健、极灵敏的电化学发光（ECL）胰岛素测定法。通过Z'值为0.82 +/- 0.03和信号背景（S/B）比为133判断，试验性能表明这是一种耐用且可靠的试验。批内和批间变异小于5%。在ECL测定中，胰岛素检测的动态范围为5 pg至5 ng。当不同体积的血清中加入外源性胰岛素时，胰岛素的回收率约为100%。这些结果表明，ECL胰岛素测定法是一种非常灵敏、稳健、非放射性的均相测定法，可成功地用于测定啮齿动物血清样品中的胰岛素水平。
In diabetes research, mouse and rat models are used for in vivo experiments, and quantification of insulin in serum samples under different pathophysiological conditions and after treatment with compounds is essential. There are few commercial radioimmunoassay and enzyme-linked immunosorbent assay (ELISA) assay kits to determine the rat/mouse plasma levels of insulin. However, reliability in insulin measurements using the available biological assays is a great concern. The authors report a robust, extremely sensitive electrochemiluminescence (ECL) insulin assay using the Origen technology platform. The assay performance, as judged by the Z' value of 0.82 +/- 0.03 and the signal-to-background (S/B) ratio of 133, suggests that this is a robust and reliable assay. The intra-assay and interassay variation is less than 5%. The dynamic range of detection for insulin is 5 pg to 5 ng in the ECL assays. Recovery of insulin was about 100% when different volumes of serum were spiked with exogenous insulin. These results suggest that the ECL insulin assay is an extremely sensitive, robust, nonradioactive homogeneous assay and can be used successfully to determine the insulin levels in rodent serum samples.