In vitro HER2 protein-induced affinity dissociation of carbon nanotube-wrapped anti-HER2 aptamers for HER2 protein detection.
In vitro HER2 protein-induced affinity dissociation of carbon nanotube-wrapped anti-HER2 aptamers for HER2 protein detection.
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DOI：
10.1039/c4an01665c
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发表时间：
2015-01
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影响因子：
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通讯作者：
J. Niazi;S. Verma;S. Niazi;A. Qureshi
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文献类型：
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作者：
J. Niazi;S. Verma;S. Niazi;A. Qureshi
基于碳纳米管包裹的抗HER2单链DNA适配子的亲和解离，建立了一种新的体外检测人表皮生长因子受体2(HER2)蛋白的方法。首先，我们选择了一个抗HER2单链DNA适配子(H2)，采用指数富集法(SELEX)进行了配体的体外连续进化。然后将荧光标记的H2单链DNA紧密结合在先前与磁性微球(MBS)偶联的碳纳米管上，形成MB-CNT-H2杂化材料。这些MB-CNTs异质结构(2.8×10(8))的负载量为H2的0.025~3.125μM。在MB-CNT-H2杂交体中，HER2蛋白诱导H2解离，解离常数Kd为270 nM。与H2-to-HER2蛋白的化学计量亲和解离比约为1：1。我们的结果表明，所建立的方法可以有效地检测游离溶液或生物样品中的天然形式的疾病生物标志物，以进行准确的诊断。
A new in vitro assay was developed to detect human epidermal growth factor receptor 2 (HER2) protein, based on affinity dissociation of carbon nanotube (CNT)-wrapped anti-HER2 ssDNA aptamers. First, we selected an anti-HER2 ssDNA aptamer (H2) using an in vitro serial evolution of ligands by an exponential enrichment (SELEX) process. Then the fluorescently labelled H2 ssDNAs were tightly packed on CNTs that had previously been coupled with magnetic microbeads (MBs), forming MB-CNT-H2 hybrids. The loading capacity of these MB-CNTs heterostructures (2.8 × 10(8)) was determined to be 0.025 to 3.125 μM of H2. HER2 protein-induced H2 dissociation occurred from MB-CNT-H2 hybrids, which was specifically induced by the target HER2 protein, with a dissociation constant (Kd) of 270 nM. The stoichiometric affinity dissociation ratio with respect to H2-to-HER2 protein was shown to be approximately 1 : 1. Our results demonstrated that the developed assay can be an effective approach in detecting native forms of disease biomarkers in free solutions or in biological samples, for accurate diagnosis.