Ultrasensitive Detection of Low-Abundance Protein Biomarkers by Mass Spectrometry Signal Amplification Assay
Ultrasensitive Detection of Low-Abundance Protein Biomarkers by Mass Spectrometry Signal Amplification Assay
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DOI：
10.1021/acs.analchem.6b01063
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发表时间：
2016-07-05
影响因子：
7.4
通讯作者：
Liu, Baohong
中科院分区：
文献类型：
--
作者：
Du, Ruijun;Zhu, Lina;Liu, Baohong
建立了一种利用金纳米粒子（AuNPs）上的标记分子进行低丰度蛋白质生物标志物超灵敏和选择性检测的质谱信号扩增方法。EpCAM和凝血酶作为模型靶点被固定在aunp上的特定适配体捕获。采用激光解吸/电离飞行时间质谱法（LDI-TOF MS）检测质量标签分子以代表蛋白质生物标志物。得益于质谱信号的放大，该方法的检测限可达100am。进一步将该方法应用于检测胎牛血清中的凝血酶和细胞裂解液中的EpCAM，以证明其在复杂生物样品中的选择性和可行性。该方案具有高灵敏度和特异性，为单细胞分析和早期疾病诊断提供了新的途径。
A mass spectrometry signal amplification method is developed for the ultrasensitive and selective detection of low abundance protein biomarkers by utilizing tag molecules on gold nanopartides (AuNPs). EpCAM and thrombin as model targets are captured by specific aptamers immobilized on the AuNPs. With laser desorption/ionization time-of-flight mass spectrometry (LDI-TOF MS), the mass tag molecules are detected to represent the protein biomarkers. Benefiting from the MS signal amplification, the assay can achieve a limit of detection of 100 aM. The method is further applied to detect thrombin in fetal bovine serum and EpCAM in cell lysates to demonstrate its selectivity and feasibility in complex biological samples. With the high sensitivity and specificity, the protocol shows great promise for providing a new route to single-cell analysis and early disease diagnosis.