Department of Chemistry, National Taiwan University, Taipei 10617, Taiwan.
Department of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung 20224, Taiwan; Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung 20224, Taiwan; School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
Biosens Bioelectron. 2016 Jan 15;75:181-7. doi: 10.1016/j.bios.2015.08.049. Epub 2015 Aug 22.
A self-assembled nanocomposite is prepared from an aqueous mixture of aptamer-modified gold nanoparticles (Apt-Au NPs), bismuth ions and chloride ions. The Apt-Au NPs are immobilized on bismuth oxychloride (BiOCl) nanosheets in situ to form Apt-Au NPs/BiOCl nanocomposites. The as-prepared nanocomposites exhibit high peroxidase-like activity for the catalytic conversion of Amplex Red (AR) to fluorescent resorufin in the presence of H2O2. The catalytic activity of Apt-Au NPs/BiOCl nanocomposites is at least 90-fold higher than that of Apt-Au NPs or BiOCl nanosheets, revealing synergistic effects on their activity. The catalytic activity of Apt-Au NPs/BiOCl nanocomposites is suppressed by vascular endothelial growth factor-A165 (VEGF-A165) molecules that specifically interact with the aptamer units (Del-5-1 and v7t-1) on the nanocomposite surface. The AR/H2O2-Apt-Au NPs/BiOCl nanocomposites probe shows high selectivity (>1000-fold over other proteins) and sensitivity (detection limit ~0.5nM) for the detection of VEGF-A165. Furthermore, the probe is employed for the detection of VEGF isoforms and for the study of interactions between VEGF and VEGF receptors. The practicality of this simple, rapid, cost-effective probe is validated by the analysis of VEGF-A165 in cell culture media, showing its great potential for the analysis of VEGF in biological samples.
一种自组装的纳米复合材料是由适配体修饰的金纳米粒子(Apt-Au NPs)、铋离子和氯离子的水溶液制备的。Apt-Au NPs 原位固定在 BiOCl 纳米片上,形成 Apt-Au NPs/BiOCl 纳米复合材料。所制备的纳米复合材料在 H2O2 存在下对 Amplex Red(AR)的催化转化具有高过氧化物酶样活性,生成荧光 Resorufin。Apt-Au NPs/BiOCl 纳米复合材料的催化活性比 Apt-Au NPs 或 BiOCl 纳米片至少高 90 倍,显示出对其活性的协同作用。血管内皮生长因子-A165(VEGF-A165)分子特异性与纳米复合材料表面上的适配体单元(Del-5-1 和 v7t-1)相互作用,抑制了 Apt-Au NPs/BiOCl 纳米复合材料的催化活性。AR/H2O2-Apt-Au NPs/BiOCl 纳米复合材料探针对 VEGF-A165 的检测具有高选择性(比其他蛋白质高 1000 倍以上)和高灵敏度(检测限约为 0.5nM)。此外,该探针还用于检测 VEGF 同工型和研究 VEGF 与 VEGF 受体之间的相互作用。该简单、快速、具有成本效益的探针的实用性通过细胞培养介质中 VEGF-A165 的分析得到验证,表明其在生物样品中分析 VEGF 具有巨大的潜力。