Ultrasensitive immunosensor for cancer biomarker proteins using gold nanoparticle film electrodes and multienzyme-particle amplification.
Ultrasensitive immunosensor for cancer biomarker proteins using gold nanoparticle film electrodes and multienzyme-particle amplification.
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DOI：
10.1021/nn800863w
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发表时间：
2009-03-24
期刊：
影响因子：
17.1
通讯作者：
Rusling, James F.
中科院分区：
文献类型：
--
作者：
Mani, Vigneshwaran;Chikkaveeraiah, Bhaskara V.;Patel, Vyomesh;Gutkind, J. Silvio;Rusling, James F.
一个密集包装的金纳米粒子平台结合多酶标记的检测抗体-磁珠生物缀合物被用作超灵敏电化学免疫传感器的基础，以检测血清中的癌症生物标志物。通过合成含有7500个辣根过氧化物酶（HRP）标记的磁性生物缀合物颗粒沿着连接到1 μm直径磁珠上的活化羧基的检测抗体（Ab 2），灵敏度大大提高。这些传感器的灵敏度为31.5 µA mL ng−1，10 µL未稀释血清中前列腺特异性抗原（PSA）的检测限（DL）为0.5 pg mL−1。这代表了5 fg PSA的超低质量DL，比先前报道的碳纳米管（CNT）森林免疫传感器好8倍，该传感器在碳纳米管上具有多个标记，并且接近或低于大多数癌症生物标志物的正常血清水平。PSA在细胞裂解物和癌症患者的人血清中的测量与标准ELISA测定具有良好的相关性。这些容易制造的金纳米粒子免疫传感器显示出良好的前景，为未来的生物电子阵列的制造。
A densely packed gold nanoparticle platform combined with a multiple-enzyme labeled detection antibody-magnetic bead bioconjugate was used as the basis for an ultrasensitive electrochemical immunosensor to detect cancer biomarkers in serum. Sensitivity was greatly amplified by synthesizing magnetic bioconjugates particles containing 7500 horseradish peroxidase (HRP) labels along with detection antibodies (Ab2) attached to activated carboxyl groups on 1 µm diameter magnetic beads. These sensors had sensitivity of 31.5 µA mL ng−1 and detection limit (DL) of 0.5 pg mL−1 for prostate specific antigen (PSA) in 10 µL of undiluted serum. This represents an ultralow mass DL of 5 fg PSA, eight fold better than a previously reported carbon nanotube (CNT) forest immunosensor featuring multiple labels on carbon nanotubes, and near or below the normal serum levels of most cancer biomarkers. Measurements of PSA in cell lysates and human serum of cancer patients gave excellent correlations with standard ELISA assays. These easily fabricated AuNP immunosensors show excellent promise for future fabrication of bioelectronic arrays.
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