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Determination of hypoxia-inducible factor-1 by using a ratiometric colorimetric test based on click-mediated growth of gold nanoparticles
Microchimica Acta ( IF 5.7 ) Pub Date : 2018-09-12 , DOI: 10.1007/s00604-018-2992-2
Hong Chen , Yan Sun , Yifei Li , Jing Zhao , Ya Cao

AbstractThe authors describe a significantly improved colorimetric nanoprobe for the determination of transcription factors (TFs). It is making use of click-mediated growth of gold nanoparticles (AuNPs) to amplify the signal-to-noise ratio. Hypoxia-inducible factor-1 (HIF-1) is an important TF that acts as a mediator of cell response to hypoxia. So, the detection of HIF-1 was chosen as the model analyte. Specifically, target HIF-1 is designed to bind to the hypoxia response element within DNA duplex. The click chemistry between the DNA duplex and alkynyl-functionalized AuNPs (AF-AuNPs) is then inhibited because of significant steric hindrance. As a result, the AF-AuNPs grow into larger-sized highly-aggregated irregular nanostructures, which in turn enable colorimetric determination. The ratio of absorbances at 620 and 560 nm increases in the 0.5 to 10 nM HIF-1 concentration range, and the detection limit is 0.27 nM. This is better by a factor of 100 than that of aggregation-based colorimetric assays. The nanoprobe is selective and can be used in complex samples. Conceivably, it may also be extended to the determination of other TFs by simply changing the used DNA duplex. Graphical abstractSchematic of a nanoprobe for detecting hypoxia-inducible factor-1 (HIF-1). Three concepts are involved: the binding of HIF-1 and hypoxia response element, the Cu+-catalyzed click chemistry between P1/P2 duplex and alkynyl-functionalized AuNPs (AF-AuNPs), and the AuNPs growth with hydroxylamine and HAuCl4.

中文翻译:

基于点击介导的金纳米粒子生长的比例比色法测定缺氧诱导因子-1

摘要作者描述了一种用于测定转录因子 (TF) 的显着改进的比色纳米探针。它利用点击介导的金纳米粒子 (AuNPs) 生长来放大信噪比。缺氧诱导因子-1 (HIF-1) 是一种重要的 TF,可作为细胞对缺氧反应的介质。因此,选择 HIF-1 的检测作为模型分析物。具体而言,目标 HIF-1 旨在与 DNA 双链体内的缺氧反应元件结合。由于显着的空间位阻,DNA 双链体和炔基功能化 AuNPs (AF-AuNPs) 之间的点击化学随后被抑制。结果,AF-AuNPs 长成更大尺寸的高度聚集的不规则纳米结构,从而能够进行比色测定。620 和 560 nm 处的吸光度比在 0. 5 至 10 nM HIF-1 浓度范围,检测限为 0.27 nM。这比基于聚合的比色测定好 100 倍。纳米探针具有选择性,可用于复杂样品。可以想象,它也可以通过简单地改变使用的 DNA 双链体来扩展到其他 TF 的确定。用于检测缺氧诱导因子-1 (HIF-1) 的纳米探针的图形摘要示意图。涉及三个概念:HIF-1 和缺氧反应元件的结合、P1/P2 双链体和炔基功能化 AuNPs(AF-AuNPs)之间的 Cu+催化点击化学,以及 AuNPs 与羟胺和 HAuCl4 的生长。纳米探针具有选择性,可用于复杂样品。可以想象,它也可以通过简单地改变使用的 DNA 双链体来扩展到其他 TF 的确定。用于检测缺氧诱导因子-1 (HIF-1) 的纳米探针的图形摘要示意图。涉及三个概念:HIF-1 和缺氧反应元件的结合、P1/P2 双链体和炔基功能化 AuNPs(AF-AuNPs)之间的 Cu+催化点击化学,以及 AuNPs 与羟胺和 HAuCl4 的生长。纳米探针具有选择性,可用于复杂样品。可以想象,它也可以通过简单地改变使用的 DNA 双链体来扩展到其他 TF 的确定。用于检测缺氧诱导因子-1 (HIF-1) 的纳米探针的图形摘要示意图。涉及三个概念:HIF-1 和缺氧反应元件的结合、P1/P2 双链体和炔基功能化 AuNPs(AF-AuNPs)之间的 Cu+催化点击化学,以及 AuNPs 与羟胺和 HAuCl4 的生长。
更新日期:2018-09-12
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