Department of Chemistry, Illinois Institute of Technology, 3101 S Dearborn St, Chicago, IL, 60616, USA.
Anal Bioanal Chem. 2018 Sep;410(24):6177-6185. doi: 10.1007/s00216-018-1224-2. Epub 2018 Jul 2.
HIV-1 protease is essential for the life cycle of the human immunodeficiency virus (HIV), and is one of the most important clinical targets for antiretroviral therapies. In this work, we developed a graphene oxide (GO)-based fluorescence biosensing platform for the rapid, sensitive, and accurate detection of HIV-1 protease, in which fluorescent-labeled HIV-1 protease substrate peptide molecules were covalently linked to GO. In the absence of HIV-1 protease, fluorescein was effectively quenched by GO. In contrast, in the presence of HIV-1 protease, it would cleave the substrate peptide into short fragments, thus producing fluorescence. Based on this sensing strategy, HIV-1 protease could be detected at as low as 1.18 ng/mL. More importantly, the sensor could successfully detect HIV-1 protease in human serum. Such GO-based fluorescent sensors may find useful applications in many fields, including diagnosis of protease-related diseases, as well as sensitive and high-throughput screening of drug candidates. Graphical abstract ᅟ.
HIV-1 蛋白酶对于人类免疫缺陷病毒(HIV)的生命周期至关重要,是抗逆转录病毒疗法的最重要的临床靶标之一。在这项工作中,我们开发了一种基于氧化石墨烯(GO)的荧光生物传感平台,用于快速、灵敏、准确地检测 HIV-1 蛋白酶,其中荧光标记的 HIV-1 蛋白酶底物肽分子通过共价键连接到 GO 上。在不存在 HIV-1 蛋白酶的情况下,GO 有效地猝灭了荧光素。相比之下,在存在 HIV-1 蛋白酶的情况下,它会将底物肽切割成短片段,从而产生荧光。基于这种传感策略,HIV-1 蛋白酶可以低至 1.18ng/mL 的浓度被检测到。更重要的是,该传感器可以成功地检测人血清中的 HIV-1 蛋白酶。这种基于 GO 的荧光传感器可能在许多领域找到有用的应用,包括蛋白酶相关疾病的诊断,以及药物候选物的灵敏和高通量筛选。