Zhang Shusheng, Yang Jun, Lin Jiehua
Key Laboratory of Eco-Biochemical Engineering, Education Ministry of China, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
Bioelectrochemistry. 2008 Feb;72(1):47-52. doi: 10.1016/j.bioelechem.2007.11.011. Epub 2007 Dec 3.
A new voltammetric enzyme-linked immunoassay system of 3,3'-diaminobenzidine (DAB)-H2O2-horseradish peroxidase (HRP) has been presented and used for the sensitive detection of carcinoembryonic antigen (CEA) in human serum. In this proposed procedure, DAB was firstly used as the electroactive substrate in the HRP catalyzed oxidation reaction in the present of H2O2. The generated product produced a sensitive second-order derivative linear sweep voltammetric peak at potential of -0.62 V (vs. SCE) in Britton-Robinson (BR) buffer solution. The free HRP could be measured in a linear range from 2.5 x 10(-6)-2.5 x 10(-2) unit/ml and a detection limit of about 1.5 x 10(-6) unit/ml. Under the optimal experiment conditions, CEA could be detected in the linear range from 0.50 to 80 ng/ml with a detection limit of 0.5 ng/ml. The proposed electrochemical enzyme-linked immunosorbent assay method is simple, inexpensive, reproducible and sensitive, which shows promising for detecting CEA in the clinical diagnosis.
一种新的3,3'-二氨基联苯胺（DAB）-H2O2-辣根过氧化物酶（HRP）伏安酶联免疫分析系统已被提出，并用于人血清中癌胚抗原（CEA）的灵敏检测。在该方法中，DAB首先在H2O2存在下作为HRP催化氧化反应中的电活性底物。生成的产物在Britton-Robinson（BR）缓冲溶液中于-0.62 V（vs. SCE）电位处产生一个灵敏的二阶导数线性扫描伏安峰。游离HRP的检测线性范围为2.5×10(-6)-2.5×10(-2)单位/毫升，检测限约为1.5×10(-6)单位/毫升。在最佳实验条件下，CEA的检测线性范围为0.50至80 ng/ml，检测限为0.5 ng/ml。所提出的电化学酶联免疫吸附测定方法简单、廉价、可重复且灵敏，在临床诊断中检测CEA显示出良好的前景。
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