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    基于免疫磁分离和酶催化放大的化学发光免疫传感器测定人血清中癌胚抗原的含量

    Determination of Carcinoembryonic Antigen in Human Serum by Chemiluminescence Immunosensor Based on Immunomagnetic Separation and Enzymatic Amplification

    • 摘要: 基于免疫磁分离和酶催化放大策略,构建了一种用于测定人血清中癌胚肮原(CEA)含量的化学发光免疫传感器。利用抗原-抗体的特异性结合,将磁球作为免疫反应的固相载体去捕获癌胚抗原(CEA),并通过在免疫磁球表面负载辣根过氧化酶(HRP)作为检测标记物,形成磁性免疫夹心复合物。通过HRP对过氧化氢的催化作用,氧化鲁米诺产生较强的化学发光信号,根据化学发光强度来定量CEA。结果显示:CEA的质量浓度在0.05~80.00 μg·L-1内与其对应的化学发光强度呈线性关系,检出限为4.5 ng·L-1;对样品进行回收试验,回收率为97.0%~100%。与其他固相载体和化学发光免疫传感器相比,该方法检测CEA的化学发光强度更强,并且具有更宽的线性范围和更低的检出限。

       

      Abstract: A chemiluminescence immunosensor for the determination of carcinoembryonic antigen (CEA) in human serum was developed based on the strategy of immunomagnetic separation and enzymatic amplitication. Based on the specific binding of antigen-antibody, magnetic balls were used as solid-phase carrier to capture CEA, and an immune complex was formed by loading horseradish peroxidase (HRP) on the surface of immunomagnetic balls as detection marker. HRP could catalyze H2O2 to oxidize luminol to generate strong chemiluminescence intensity, and CEA was quantified according to the chemiluminescence intensity. As shown by the results, linear relationship between value of mass concentration of CEA and its chemiluminescence intensity was kept in the range of 0.05-80.00 μg·L-1, with detection limit of 4.5 ng·L-1. Test for recovery was made on the samples, giving results in the range of 97.0%-100%. Compared with other solid phase carriers and chemiluminescence immunosensors, the chemiluminescence intensity of CEA detected by this method was stronger, and it had the wider linear range and lower detection limit.

       

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