Key Laboratory of Luminescence and Real-Time Analysis of the Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing 400715, China.
Biosens Bioelectron. 2013 Sep 15;47:445-50. doi: 10.1016/j.bios.2013.03.030. Epub 2013 Mar 21.
Graphene oxide (GO) has been widely used to develop fluorescence resonance energy transfer (FRET) biosensors for tumor markers (e.g., matrix metalloproteinases, MMPs) due to its superior fluorescence quenching capacity and unique adsorption characteristics for biomolecules. In this study, fluorescein isothiocyanate-labeled peptide (Pep-FITC) was assembled onto the GO surface through covalent binding to construct a GO-Pep-FITC FRET sensor for sensitive, rapid, and accurate detection of MMP-2 in complex serum samples. Compared to similar GO-based FRET sensors fabricated through physical adsorption, the as prepared ones via covalent binding are significantly more stable under physiological conditions, enabling their detection of MMP-2 with high sensitivity (detection limit: 2.5ng/mL). More importantly, it allows for rapid MMP-2 detection (within 3h) even in complex biological samples with satisfactory accuracy and the relative standard deviation ≤7.03%. Our studies further suggest that such a platform developed here for sensitive, rapid, and accurate detection of biomarkers holds great promise for clinical diagnosis of protease-related diseases.
氧化石墨烯（GO）因其优异的荧光猝灭能力和独特的生物分子吸附特性，已被广泛用于开发用于肿瘤标志物（如基质金属蛋白酶，MMPs）的荧光共振能量转移（FRET）生物传感器。在这项研究中，通过共价键合将异硫氰酸荧光素标记的肽（Pep-FITC）组装到 GO 表面上，构建了用于敏感，快速和准确检测复杂血清样品中 MMP-2 的 GO-Pep-FITC FRET 传感器。与通过物理吸附制备的类似基于 GO 的 FRET 传感器相比，通过共价键合制备的传感器在生理条件下更加稳定，使其能够以高灵敏度（检测限：2.5ng/mL）检测 MMP-2。更重要的是，即使在复杂的生物样品中，它也可以快速（在 3 小时内）检测 MMP-2，准确性令人满意，相对标准偏差≤7.03％。我们的研究进一步表明，该平台具有用于敏感，快速和准确检测生物标志物的巨大潜力，可用于与蛋白酶相关的疾病的临床诊断。
