基于巯基丁二酰肼铜(Ⅱ)单层修饰电极的HIV-p24电化学免疫传感器

Electrochemical Immunosensor for Human Immunodeficiency Virus p24 Antigen Based on Mercapto Succinic Acid Hydrazide Copper Monolayer Modified Gold Electrode

研制了基于巯基丁二酰肼铜(Ⅱ)(CuL)单层修饰金电极(Au|CuL)测定艾滋病人血清中HIV核心抗原p24水平的免疫传感器。该传感器通过将CuL直接自组装到金电极表面制备而成。CuL对H2O2的还原具有催化作用,可作为p24抗体(antip24)上标记的辣根过氧化物酶(HRP)与电极之间电子传递媒介体。结合竞争性免疫分析,HRP-antip24抗体(HRP-antip24)与待测p24反应生成的免疫结合物游离在反应池中,通过测定在免疫结合物上HRP作用下电极表面CuL对H2O2催化增强电流,采用示差脉冲伏安法(DPV)直接获得待测抗原的含量。该免疫传感器的测定无需分离和洗涤步骤,温育时间短,为艾滋病诊断提供了一种新的方法。

An immunosensor based on mercapto succinic acid hydrazide copper（ Ⅱ ） （CuL） monolayer modi- fied gold electrode（AuICuL） was proposed for the determination of human immuno deficiency virus p24 anti- gen in acquired immuno deficiency syndrome （AIDs） patients＇ serum. CuL displays reductive electrocatalyza- tion for H2O2 and can be employed as electron transfer media between HRP and electrode. Based on a compe- tition immuno-reaction a differential pulse voltammetric （DPV） determination method for p24 antigen was established by the peak current increase of the immobilized CuL. The current increase results from the enhanced catalytic current of H2O2 reduction with the help of HRP labeled immuno-compound. The free immuno-compound was produced by competitive binding to the limited amount of HRP labeled p24 antibody （HRP-anti p24） from the p24 in sample solution and immobilized on the bottom of 96-well plates. The pro- posed method needs neither separation nor washing up step, which simplifies the immunoassay procedure, shortens the analytical time, and provides a new promising platform for diagnosing the AIDs disease.