Department of Pharmaceutical Sciences and Technology, Birla Institute of Technology, Mesra, Ranchi, Jharkhand, 835215, India.
Department of Chemistry, University of Miami, Coral Gables, FL, 33146, USA.
Sci Rep. 2023 Sep 4;13(1):14560. doi: 10.1038/s41598-023-41885-w.
Frequent mutation and variable immunological protection against vaccination is a common feature for COVID-19 pandemic. Early detection and confinement remain key to controlling further spread of infection. In response, we have developed an aptamer-based system that possesses both diagnostic and therapeutic potential towards the virus. A random aptamer library (~ 10 molecules) was screened using systematic evolution of ligands by exponential enrichment (SELEX) and aptamer R was identified as a potent binder for the SARS-CoV-2 spike receptor binding domain (RBD) using in vitro binding assay. Using a pseudotyped viral entry assay we have shown that aptamer R specifically inhibited the entry of a SARS-CoV-2 pseudotyped virus in HEK293T-ACE2 cells but did not inhibit the entry of a Vesicular Stomatitis Virus (VSV) glycoprotein (G) pseudotyped virus, hence establishing its specificity towards SARS-CoV-2 spike protein. The antiviral potential of aptamers R and J (same central sequence as R but lacking flanked primer regions) was tested and showed 95.4% and 82.5% inhibition, respectively, against the SARS-CoV-2 virus. Finally, intermolecular interactions between the aptamers and the RBD domain were analyzed using in silico docking and molecular dynamics simulations that provided additional insight into the binding and inhibitory action of aptamers R and J.
频繁的突变和针对疫苗接种的可变免疫保护是 COVID-19 大流行的共同特征。早期检测和隔离仍然是控制感染进一步传播的关键。有鉴于此，我们开发了一种基于适体的系统，该系统具有针对该病毒的诊断和治疗潜力。使用指数富集的配体系统进化（SELEX）筛选了一个随机适体文库（约 10 个分子），并通过体外结合试验鉴定出适体 R 是 SARS-CoV-2 刺突受体结合域（RBD）的有效结合物。使用假型病毒进入试验，我们已经表明适体 R 特异性抑制了 SARS-CoV-2 假型病毒在 HEK293T-ACE2 细胞中的进入，但不抑制水疱性口炎病毒（VSV）糖蛋白（G）假型病毒的进入，因此确立了其对 SARS-CoV-2 刺突蛋白的特异性。测试了适体 R 和 J（与 R 具有相同的中心序列但缺少侧翼引物区域）的抗病毒潜力，发现它们对 SARS-CoV-2 病毒的抑制率分别为 95.4%和 82.5%。最后，使用计算对接和分子动力学模拟分析了适体与 RBD 结构域之间的分子间相互作用，这为适体 R 和 J 的结合和抑制作用提供了更多的见解。
