Gold nanoparticle-labeled detection antibodies for use in an enhanced electrochemical immunoassay of hepatitis B surface antigen in human serum
Gold nanoparticle-labeled detection antibodies for use in an enhanced electrochemical immunoassay of hepatitis B surface antigen in human serum
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DOI：
10.1007/s00604-009-0184-9
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发表时间：
2009-07
影响因子：
5.7
通讯作者：
Sheng-Han Wu;Z. Zhong;Dong Wang;Mengxia Li;Y. Qing;N. Dai;Zeng-peng Li
中科院分区：
文献类型：
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作者：
Sheng-Han Wu;Z. Zhong;Dong Wang;Mengxia Li;Y. Qing;N. Dai;Zeng-peng Li
基于金纳米颗粒的电化学免疫分析在检测B型肝炎表面抗原（HBsAg）方面的应用已得到广泛研究，但大多数情况下，它们表现出较低的灵敏度。我们描述了一种新的电化学免疫分析的抗原-抗体反应的信号放大与纳米金为基础的生物条形码技术相结合的制作。首先将B型肝炎表面抗体（HBsAg Ab）固定在纳米金/硫代巴比妥/DNA修饰的金电极上，然后使用纳米金编码的辣根过氧化物酶-HBsAg Ab偶联物作为二抗，采用ELISA型免疫测定格式检测HBsAg。在最佳条件下，该免疫复合物对H2 O2体系的电流响应与HBsAg浓度在0.5 ~ 650 ng·mL-1范围内呈良好的线性关系，检出限为0.1 ng·mL-1（S/N = 3）。该免疫传感器具有良好的精密度、重现性和稳定性。随后，免疫传感器用于检测人血清标本中的HBsAg。分析结果与标准化学发光酶联免疫吸附试验所获得的结果一致。
The application of gold nanoparticle-based electrochemical immunoassays have been extensively studied for the detection of hepatitis B surface antigen (HBsAg), but most often they exhibit low sensitivity. We describe the fabrication of a new electrochemical immunoassay for signal amplification of the antigen-antibody reaction combined with the nanogold-based bio-barcode technique. Hepatitis B surface antibody (HBsAb) was initially immobilized on a nanogold/thionine/DNA-modified gold electrode, and then a sandwich-type immunoassay format was employed for the detection of HBsAg using nanogold-codified horseradish peroxidase-HBsAb conjugates as secondary antibodies. Under optimal conditions, the current response of the sandwich-type immunocomplex relative to the H2O2system was proportional to HBsAg concentration in the range from 0.5 to 650 ng·mL−1with a detection limit of 0.1 ng·mL−1(S/N = 3). The precision, reproducibility and stability of the immunosensor were acceptable. Subsequently, the immunosensors were used to assay HBsAg in human serum specimens. Analytical results were in agreement with those obtained by the standard chemiluminescence enzyme-linked immunosorbent assay.