A competitive immunochromatographic assay for testosterone based on electrochemical detection.
A competitive immunochromatographic assay for testosterone based on electrochemical detection.
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DOI：
10.1016/j.talanta.2007.05.008
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发表时间：
2007-10
期刊：
影响因子：
6.1
通讯作者：
Kumi Y. Inoue;Pascal Ferrante;Y. Hirano;T. Yasukawa;H. Shiku;T. Matsue
中科院分区：
文献类型：
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作者：
Kumi Y. Inoue;Pascal Ferrante;Y. Hirano;T. Yasukawa;H. Shiku;T. Matsue
使用硝酸纤维素膜的免疫色谱分析与使用电极芯片的电化学检测相结合，以定量检测睾酮作为模型分析物。电极芯片由金工作电极、对电极和伪参比电极组成，均制作在3.2mm×3.2mm的孔底。通过使含有睾酮和辣根过氧化物酶（HRP）标记的睾酮（竞争对手）的溶液流过膜来启动膜上的竞争性免疫反应。将制得的膜置于含有二茂铁甲醇（FcOH）和H2 O2的溶液中，置于电极芯片的孔中，用安培法检测酶反应。捕获在膜上的标记HRP催化FcOH氧化成氧化形式FcOH+，其通过电极芯片电化学还原。在1- 625 ng/ml范围内，还原电流随睾酮浓度的增加而减小。
An immunochromatographic assay using nitrocellulose membrane was combined with electrochemical detection using an electrode chip in order to quantitatively detect testosterone as a model analyte. The electrode chip consisted of a gold working electrode, a counter electrode and a pseudo-reference electrode, all fabricated on the bottom of a 3.2mm×3.2mm well. Competitive immunoreactions on the membrane were initiated by flowing a solution containing testosterone and horseradish peroxidase (HRP)-labeled testosterone (a competitor) over the membrane. Prepared membrane was placed in a solution containing ferrocenemethanol (FcOH) and H2O2in the well of the electrode chip, and the enzyme reaction was detected by amperometry. Labeled HRP captured on the membrane catalyzed the oxidation of FcOH to the oxidized form FcOH+, which was reduced electrochemically by the electrode chip. The electrochemical response of the reduction current decreased with increasing concentration of testosterone over the range 1–625ng/ml.