Aptamer-modified gold nanoparticles for colorimetric determination of platelet-derived growth factors and their receptors
Aptamer-modified gold nanoparticles for colorimetric determination of platelet-derived growth factors and their receptors
复制标题
DOI：
10.1021/ac050957q
复制
发表时间：
2005-09-01
影响因子：
7.4
通讯作者：
Chang, HT
中科院分区：
文献类型：
--
作者：
Huang, CC;Huang, YF;Chang, HT
我们已经开发了一种高度特异性的传感系统，用于血小板衍生生长因子（PDGF）和血小板衍生生长因子受体（PDGFR），使用金纳米粒子（GNP）。我们合成了用对PDGF特异性的适体（Apt-GNP）修饰的GNP，并通过监测由于聚集而发生的Apt-GNP的颜色和消光的变化来使用它们检测PDGF。Apt-GNP的颜色在低浓度（400 nM）下从红色变为紫色。我们发现Apt-GNP对三种PDGF的敏感性是高度盐依赖性的，最佳条件为200 mM NaCl。当绘制不同Apt-GNPs浓度下Apt-GNPs在650和530 nm处的消光系数与PDGF-AA浓度的比值时，我们获得了双相曲线。对于0.42 nM Apt-GNP，该消光比的增加和减少的线性范围分别为2.5-10和10-20 nM，对于8.4 nM Apt-GNP，分别为25-75和75-200 nM。当使用8.4 nM Apt-GNP时，PDGF-AB的消光比增加和减少的相应线性范围分别为15-100和100-400 nM，PDGF-BB的消光比增加和减少的相应线性范围分别为35 - 150和150-400 nM。此外，我们已经开发了一种均相测定法，以检测浓度低至3.2 nM的PDGF受体-β（PDGFR-β），基于Apt-GNP和PDGFR-β之间对PDGF-BB的竞争。我们在本文中提出的结果意味着Apt-GNP在蛋白质分析和癌症诊断中有实际应用。
We have developed a highly specific sensing system for platelet-derived growth factors (PDGFs) and platelet-derived growth factor receptors (PDGFR) that uses gold nanoparticles (GNPs). We synthesized GNPs modified with an aptamer (Apt-GNPs) that is specific to PDGFs and used them to detect PDGFs by monitoring the changes in the color and extinction of the Apt-GNPs that occur as a result of aggregation. The color of the Apt-GNPs changes from red to purple at low concentrations ( 400 nM). We found that the sensitivity of the Apt-GNPs for the three PDGFs is highly salt-dependent, with an optimum condition of 200 mM NaCl. We obtained biphasic curves when plotting of the ratios of the extinction coefficients of the Apt-GNPs at 650 and 530 nm against the concentrations of PDGF-AA at various concentrations of Apt-GNPs. The linear ranges of the increases and decreases in this extinction ratio are 2.5-10 and 10-20 nM, respectively, for 0.42 nM Apt-GNPs and 25-75 and 75-200 nM, respectively, for 8.4 nM Apt-GNPs. When using 8.4 nM Apt-GNPs, the corresponding linear ranges of the increases and decreases in this extinction ratio are 15-100 and 100-400 nM, respectively, for PDGF-AB and 35150 and 150-400 nM, respectively, for PDGF-BB. In addition, we have developed a homogeneous assay to detect the PDGF receptor-beta (PDGFR-beta) at concentrations as low as 3.2 nM, on the basis of the competition between the Apt-GNPs and PDGFR-beta for PDGF-BB. The results we present in this paper imply that there are practical applications of Apt-GNPs in protein analysis and cancer diagnosis.