Electrochemical immunoassay for the protein biomarker mucin 1 and for MCF-7 cancer cells based on signal enhancement by silver nanoclusters
Electrochemical immunoassay for the protein biomarker mucin 1 and for MCF-7 cancer cells based on signal enhancement by silver nanoclusters
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基于银纳米簇信号增强的蛋白质生物标志物粘蛋白 1 和 MCF-7 癌细胞的电化学免疫分析
DOI：
10.1007/s00604-015-1471-2
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发表时间：
2015-06-01
影响因子：
5.7
通讯作者：
Yang, Minghui
中科院分区：
文献类型：
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作者：
Guo, Qingjun;Li, Xiangzhi;Yang, Minghui
描述了用于检测蛋白质生物标志物粘蛋白1（MUC-1）和其中MUC-1过表达的MCF-7型乳腺癌细胞的电化学免疫测定。该方法是基于使用的银纳米簇（银-NC）作为信号探针。在序列为5 '-GCAGTTGATCCTTTGGATACCTGG-C-12- 3'的DNA链存在下，通过化学还原合成Ag-NC。该链含有粘蛋白1适体（GCAGTTGATCCTTTGGATACCCTGG），可特异性结合MUC 1和模板（C-12），用于合成Ag-NC。该测定包括以下步骤：（1）通过将针对MUC-1的抗体固定在玻碳电极上来构建免疫传感器;（2）添加含有MUC-1的样品;（3）添加Ag-NC;（4）通过银增强过程（在Ag-NC上沉积金属银）进行信号放大;（5）通过方波伏安法进行测量。在0.11 V（vs. SCE）电位下测量的电流与MUC-1浓度在1至500 nM范围内呈对数相关，检测限为0.5 nM。我们还证明，由于细胞表面存在MUC-1蛋白，因此可以通过该方法以高灵敏度（50个细胞/mL）检测MCF-7癌细胞。
An electrochemical immunoassay is described for the detection of the protein biomarker mucin 1 (MUC-1) and of breast cancer cells of type MCF-7 where MUC-1 is overexpressed. The method is based on the use of silver nanoclusters (Ag-NCs) acting as a signalling probe. The Ag-NCs were synthesized via chemical reduction in the presence of a DNA strand with the sequence of 5'-GCAGTTGATCCTTTGGATACCCTGG-C-12-3'. The strand contains mucin 1 aptamer (GCAGTTGATCCTTTGGATACCCTGG) that can specifically bind to MUC1 and the template (C-12) for synthesis of Ag-NCs. The assay involves the following steps: (1) Construction of an immunosensor by immobilizing the antibody against MUC-1 on a glassy carbon electrode; (2) addition of sample containing MUC-1; (3) addition of Ag-NCs; (4) signal amplification via silver enhancement process (deposition of metal silver on Ag-NCs); (5) measurement via square wave voltammetry. The current measured at a potential of 0.11 V (vs. SCE) is logarithmically related to the concentration of MUC-1 in the 1 to 500 nM range, with a detection limit of 0.5 nM. We also demonstrate that MCF-7 cancer cells can be detected by this method with high sensitivity (50 cells per mL) due to the presence of MUC-1 proteins on the cell surface.