Radiation Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), 1266 Sinjeong-dong, Jeongeup, Jeonbuk 580-185, Republic of Korea.
Talanta. 2011 Oct 15;85(5):2639-42. doi: 10.1016/j.talanta.2011.08.035. Epub 2011 Aug 22.
In this study, we elucidated a small molecule inhibitor on viral protein NS5B identified through a high-throughput screening strategy using optical nanoparticle-based RNA oligonucleotide. We have previously shown that quantum dots (QDs)-RNA oligonucleotide can specifically recognize the HCV viral proteins. We have also demonstrated that conjugated QDs-RNA oligonucleotide can specifically and sensitively interact with designed biochips [1,2]. Among the flavonoids examined, (-)-epigallocatechin gallate (EGCG) demonstrated a remarkable inhibition activity on HCV viral protein, NS5B. (-)-Epigallocatechin gallate, at 0.005 μg mL(-1) or more, concentration-dependently attenuated the binding affinity on a designed biochip as evidenced by QDs-RNA oligonucleotide. At a concentration of 0.1 μg mL(-1), (-)-epigallocatechin gallate showed a 50% inhibition activity on QDs-RNA oligonucleotide biochip assay. We screened a small molecule inhibitor on the viral protein, NS5B, identified through a high-throughput screening strategy using on-chip optical nanoparticle-based RNA oligonucleotide on chip. In this designed strategy, the convenient and efficient screening and development of an on-chip viral protein inhibitor using a QDs-RNA oligonucleotide assay is achievable with high sensitivity and simplicity. In addition, this platform is expected to be applicable toward the inhibitor screening of other types of diseases.
在这项研究中，我们通过使用基于光学纳米粒子的 RNA 寡核苷酸的高通量筛选策略，阐明了一种针对病毒蛋白 NS5B 的小分子抑制剂。我们之前已经表明，量子点（QD）-RNA 寡核苷酸可以特异性识别 HCV 病毒蛋白。我们还证明了共轭 QD-RNA 寡核苷酸可以特异性和灵敏地与设计的生物芯片相互作用[1,2]。在所检查的类黄酮中，（-）-表没食子儿茶素没食子酸酯（EGCG）对 HCV 病毒蛋白 NS5B 表现出显著的抑制活性。（-）-表没食子儿茶素没食子酸酯在 0.005 μg mL（-1）或更高浓度下，浓度依赖性地减弱了设计的生物芯片上的结合亲和力，这一点可以通过 QD-RNA 寡核苷酸证明。在 0.1 μg mL（-1）的浓度下，（-）-表没食子儿茶素没食子酸酯对 QD-RNA 寡核苷酸生物芯片测定显示出 50%的抑制活性。我们通过在芯片上使用基于芯片的光学纳米粒子 RNA 寡核苷酸的高通量筛选策略，筛选出针对病毒蛋白 NS5B 的小分子抑制剂。在这个设计的策略中，使用 QD-RNA 寡核苷酸测定可以方便、有效地筛选和开发针对病毒蛋白的抑制剂，具有高灵敏度和简单性。此外，该平台有望应用于其他类型疾病的抑制剂筛选。
