Capillary electrophoretic enzyme immunoassay with electrochemical detection for thyroxine
Capillary electrophoretic enzyme immunoassay with electrochemical detection for thyroxine
复制标题
DOI：
10.1016/s0003-2697(02)00508-0
复制
发表时间：
2003-02-01
影响因子：
2.9
通讯作者：
Jin, WR
中科院分区：
文献类型：
--
作者：
He, ZH;Jin, WR
建立了毛细管电泳酶免疫分析-电化学检测法（CE-EIA-艾德）。在该方法中，抗原（Ag）与辣根过氧化物酶（HRP）标记的抗原（HRP-Ag）竞争有限数量的抗体（Ab）结合位点。游离的HRP-Ag和结合的HRP-Ag-Ab复合物在分离毛细管中通过毛细管电泳分离。然后，它们在反应毛细管中用H2 O2催化它们的酶底物3，3 '，5，5'-四甲基联苯TMB（还原形式）的氧化，反应毛细管在分离毛细管之后。在反应毛细管的出口处使用碳纤维微盘束电极以安培法测定反应产物TMB（氧化形式）。由于酶的扩增，TMB（Ox）的浓度远高于游离HRP-Ag和结合HRP-Ag Ab复合物的浓度。因此，CE-EIA-艾德的检测限（LOD）非常低。方法已用于人血清中甲状腺素的测定。达到的LOD浓度为3.8 x 10（-9）mol/L，相当于质量LOD为23.2 μ mol。(C)2003 Elsevier Science（美国）。All rights reserved.
A capillary electrophoretic enzyme immunoassay with electrochemical detection (CE-EIA-ED) has been developed. In this method, antigen (Ag) competes with horseradish peroxidase (HRP)-labeled antigen (HRP-Ag) for a limited number of antibody (Ab) binding sites. The free HRP-Ag and the bound HRP-Ag-Ab complex are separated by capillary electrophoresis in a separation capillary. Then they catalyze the oxidation of their enzyme substrate 3,3',5,5'-tetramethylbenzide TMB (reduced form)) with H2O2 in a reaction capillary, which follows the separation capillary. The reaction product TMB (oxidized form)) is amperometrically determined using a carbon fiber microdisk bundle electrode at the outlet of the reaction capillary. Due to the amplification of the enzyme, the concentration of TMB(Ox) is much higher than those of free HRP-Ag and the bound HRP-Ag Ab complex. Therefore, the limit of detection (LOD) of CE-EIA-ED is very low. The method has been used to determine thyroxine in human serum. A concentration of LOD of 3.8 x 10(-9) mol/L, which corresponds to a mass LOD of 23.2 mumol, was achieved. (C) 2003 Elsevier Science (USA). All rights reserved.