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基于适体/金碳点/磁性石墨烯氧化纳米片的高灵敏高尔基糖蛋白 3 检测策略作为荧光生物传感器。

A highly sensitive strategy for glypican-3 detection based on aptamer/gold carbon dots/magnetic graphene oxide nanosheets as fluorescent biosensor.

机构信息

School of Life and Environmental Sciences, Guilin University of Electronic Technology, Guilin, Guangxi, 541004, People's Republic of China.

College of Chemistry, Guangdong University of Petrochemical Technology, Guandu Road, Maoming, Guangdong, 525000, People's Republic of China.

出版信息

Anal Bioanal Chem. 2022 Sep;414(22):6441-6453. doi: 10.1007/s00216-022-04201-5. Epub 2022 Jul 5.

DOI:10.1007/s00216-022-04201-5
PMID:35788872
Abstract

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths in China. Glypican-3 (GPC3) is a specific antigen related to HCC, which is widely used in clinical detection as a reliable marker of HCC. In this paper, a highly sensitive homogeneous apatasensor was designed for GPC3 detection based on fluorescence resonance energy transfer (FRET) where the GPC3 aptamer labelled gold carbon dots (AuCDs-GPC3) are used as a donor and magnetic graphene oxide (FeO/GO) nanosheets are used as an acceptor. A one-step hydrothermal method was used to synthesize AuCDs to provide sufficient fluorescence. The FRET phenomenon exists between AuCDs-GPC3 and FeO/GO, which weakens the fluorescence intensity of the whole system. When the target GPC3 is added to the FRET system, the fluorescent AuCDs-GPC3 binds to the GPC3 and forms a folded structure, which leads to AuCDs-GPC3 separation from FeO/GO nanosheets. The FeO/GO is then magnetically separated so that the fluorescence of free labelled AuCDs-GPC3 is restored. Under the optimum conditions, the fluorescence recovery rate is linearly correlated with the concentration of GPC3 (5-100 ng·mL) and the detection limit is 3.01 ng·mL (S/N = 3). This strategy shows recoveries from 98.76 to 101.29% in real human serum samples and provides an immediate and effective detection method for the quantification of GPC3 with great potential applications for early diagnosis of HCC. A sensitive homogeneous FRET-based apatasensor was designed for GPC3 detection where the AuCDs-GPC3 is a donor and FeO/GO nanosheets are an acceptor. The GPC3 fluorescent aptasensor combines wider output range with low cost, high specificity, and good anti-interference.

摘要

肝细胞癌(HCC)是中国癌症相关死亡的第二大主要原因。磷脂酰聚糖-3(GPC3)是一种与 HCC 相关的特异性抗原,广泛应用于临床检测,作为 HCC 的可靠标志物。本文基于荧光共振能量转移(FRET)设计了一种用于 GPC3 检测的高灵敏度均相适配体传感器,其中 GPC3 适配体标记的金碳点(AuCDs-GPC3)作为供体,磁性氧化石墨烯(FeO/GO)纳米片作为受体。采用一步水热法合成 AuCDs 以提供足够的荧光。AuCDs-GPC3 与 FeO/GO 之间存在 FRET 现象,这削弱了整个系统的荧光强度。当目标 GPC3 被添加到 FRET 系统中时,荧光 AuCDs-GPC3 与 GPC3 结合形成折叠结构,导致 AuCDs-GPC3 从 FeO/GO 纳米片上分离。然后通过磁性分离 FeO/GO,从而恢复游离标记的 AuCDs-GPC3 的荧光。在最佳条件下,荧光恢复率与 GPC3 的浓度(5-100ng·mL)呈线性相关,检测限为 3.01ng·mL(S/N=3)。该策略在真实人血清样品中的回收率为 98.76%至 101.29%,为 GPC3 的定量提供了一种即时有效的检测方法,具有 HCC 早期诊断的巨大应用潜力。设计了一种基于 AuCDs-GPC3 作为供体和 FeO/GO 纳米片作为受体的用于 GPC3 检测的灵敏均相 FRET 适配体传感器。GPC3 荧光适配体传感器具有更宽的输出范围、低成本、高特异性和良好的抗干扰性。

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