Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources, Guangxi Normal University, Guilin, 541004, China.
Department of Chemistry and Pharmacy, Guilin Normal College, Guilin, 541001, Guangxi, China.
Mikrochim Acta. 2019 Feb 22;186(3):204. doi: 10.1007/s00604-019-3312-1.
The extraordinary fluorescence quenching capability of graphene oxide (GO) was coupled to the specific recognition capability of aptamers to design a four-color fluorescent nanoprobe for multiplexed detection and imaging of tumor-associated proteins in living cells. Specifically, alpha-fetoprotein (AFP), vascular endothelial growth factor-165 (VEGF), carcinoembryonic antigen (CEA), and human epidermal growth factor receptor 2 (HER2) were detected. Due to strong π interaction, the fluorescence of labeled aptamers is quenched by GO. Four fluorophore-labeled aptamers that bind the tumor-associated proteins were adsorbed on GO to form the four-color nanoprobe with quenched fluorescence. The nanoprobes were internalized into cells via endocytosis, where the aptamer/GO nanoprobes bind the intracellular tumor-associated proteins. The aptamer-protein complexes thus formed detach from GO, and fluorescence recovers. Each analyte has its typical color (AFP: blue; VEGF165: green; CEA: yellow; HER2: red). As a result, simultaneous detection and imaging of multiple tumor-associated proteins in living cells were achieved. This nanoprobe has a fast response and is highly specific and biocompatible. The linear ranges for AFP, VEGF, CEA, and HER2 are 0.8 nM-160 nM, 0.5 nM-100 nM, 1.0 nM-200 nM, and 1.2 nM-240 nM, respectively. Detection limits were 0.45 nM for AFP, 0.30 nM for VEGF, 0.62 nM for CEA, and 0.96 nM for HER2. The probe allows for a fast distinction between tumor cells and normal cells via imaging. Graphical abstract Schematic presentation of the development of a four-color fluorometic method based on aptamer and graphene oxide for simultaneous detection and imaging of alpha-fetoprotein, vascular endothelial growth factor-165, carcinoembryonic antigen and human epidermal growth factor receptor 2 in living cells.
基于适配体和氧化石墨烯的四色荧光法同时检测活细胞中α-胎蛋白、血管内皮生长因子 165、癌胚抗原和人表皮生长因子受体 2
设计了一种基于适配体和氧化石墨烯的四色荧光法，用于同时检测和成像活细胞中的α-胎蛋白（AFP）、血管内皮生长因子-165（VEGF）、癌胚抗原（CEA）和人表皮生长因子受体 2（HER2）。由于强烈的π相互作用，标记适配体的荧光被 GO 猝灭。将四种与肿瘤相关蛋白结合的荧光标记适配体吸附到 GO 上，形成荧光猝灭的四色纳米探针。纳米探针通过内吞作用进入细胞，其中适配体/GO 纳米探针与细胞内的肿瘤相关蛋白结合。形成的适配体-蛋白复合物因此从 GO 上脱离，荧光恢复。每种分析物都有其典型的颜色（AFP：蓝色；VEGF165：绿色；CEA：黄色；HER2：红色）。因此，实现了活细胞中多种肿瘤相关蛋白的同时检测和成像。该纳米探针具有快速响应、高特异性和生物相容性。AFP、VEGF、CEA 和 HER2 的线性范围分别为 0.8 nM-160 nM、0.5 nM-100 nM、1.0 nM-200 nM 和 1.2 nM-240 nM。AFP 的检测限为 0.45 nM，VEGF 的检测限为 0.30 nM，CEA 的检测限为 0.62 nM，HER2 的检测限为 0.96 nM。该探针通过成像能够快速区分肿瘤细胞和正常细胞。
Zotero 插件