Department of Endocrine and Breast Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400042, China.
Guangshan County People's Hospital, Xinyang, 465450, Henan, China.
J Nanobiotechnology. 2022 Jun 27;20(1):306. doi: 10.1186/s12951-022-01512-8.
Protein-stabilized gold nanoclusters (Prot-Au NCs) have been widely used in biosensing and cell imaging owing to their excellent optical properties and low biotoxicity. However, several Prot-Au NCs reported in the literature do not retain the biological role of the protein, which greatly limits their ability to directly detect biomarkers. This study demonstrated for the first time the successful synthesis of dual-function avidin-stabilized gold nanoclusters (Av-Au NCs) using a one-pot method. The resulting Av-Au NCs exhibited intense blue and red emissions under 374 nm excitation. Furthermore, the Av-Au NCs retained the native functionality of avidin to bind to biotin. When DNA strands modified with biotin at both ends (i.e., linker chains) were mixed with Av-Au NCs, large polymers were formed, indicating that Av-Au NCs could achieve fluorescence signal amplification by interacting with biotin. Taking advantage of the aforementioned properties, we constructed a novel enzyme-free fluorescent biosensor based on the Av-Au NCs-biotin system to detect DNA. The designed fluorescent biosensor could detect target DNA down to 0.043 nM, with a wide line range from 0.2 nM to 20 µM. Thus, these dual-functional Av-Au NCs were shown to be an excellent fluorescent material for biosensing.
基于其优异的光学性质和低生物毒性,蛋白稳定的金纳米团簇(Prot-Au NCs)已被广泛应用于生物传感和细胞成像。然而,文献中报道的几种 Prot-Au NCs 并不保留蛋白质的生物学功能,这极大地限制了它们直接检测生物标志物的能力。本研究首次成功地使用一锅法合成了具有双重功能的亲和素稳定的金纳米团簇(Av-Au NCs)。所得的 Av-Au NCs 在 374nm 激发下表现出强烈的蓝色和红色发射。此外,Av-Au NCs 保留了亲和素与生物素结合的天然功能。当两端都修饰有生物素的 DNA 链(即连接链)与 Av-Au NCs 混合时,会形成大聚合物,表明 Av-Au NCs 可以通过与生物素相互作用实现荧光信号放大。利用上述特性,我们基于 Av-Au NCs-生物素系统构建了一种新型的无酶荧光生物传感器来检测 DNA。所设计的荧光生物传感器可以检测低至 0.043nM 的靶 DNA,线性范围从 0.2nM 到 20µM 很宽。因此,这些双重功能的 Av-Au NCs 被证明是一种用于生物传感的优异荧光材料。