Department of BioNano Technology, Gachon University, 1342 Seongnamdae-ro, Sujeong-gu, Seongnam 13120, Gyeonggi, Korea.
Int J Mol Sci. 2021 Dec 29;23(1):366. doi: 10.3390/ijms23010366.
An effective strategy to detect biological thiols (biothiols), including glutathione (GSH), cysteine (Cys), and homocysteine (Hcy), holds significant incentive since they play vital roles in many cellular processes and are closely related to many diseases. Here, we demonstrated that hybrid nanoflowers composed of crystalline copper phosphate and horseradish peroxidase (HRP) served as a functional unit exhibiting dual catalytic activities of biothiol oxidase and HRP, yielding a cascade reaction system for a sensitive one-pot fluorescent detection of biothiols. The nanoflowers were synthesized through the anisotropic growth of copper phosphate petals coordinated with the amine/amide moieties of HRP, by simply incubating HRP and copper(II) sulfate for three days at room temperature. Copper phosphates within the nanoflowers oxidized target biothiols to generate HO, which activated the entrapped HRP to oxidize the employed Amplex UltraRed substrate to produce intense fluorescence. Using this strategy, biothiols were selectively and sensitively detected by monitoring the respective fluorescence intensity. This nanoflower-based strategy was also successfully employed for reliable quantification of biothiols present in human serum, demonstrating its great potential for clinical diagnostics.
一种有效检测生物硫醇（biothiols）的策略，包括谷胱甘肽（GSH）、半胱氨酸（Cys）和同型半胱氨酸（Hcy），因为它们在许多细胞过程中发挥着重要作用，并且与许多疾病密切相关，所以具有重要的激励作用。在这里，我们展示了由结晶磷酸铜和辣根过氧化物酶（HRP）组成的混合纳米花作为一个具有生物硫醇氧化酶和 HRP 双重催化活性的功能单元，产生了一个级联反应系统，用于灵敏的一步法荧光检测生物硫醇。纳米花是通过铜磷酸盐花瓣与 HRP 的胺/酰胺部分的配位进行各向异性生长合成的，只需在室温下将 HRP 和硫酸铜孵育三天。纳米花内的磷酸铜将目标生物硫醇氧化生成 HO，激活包裹的 HRP 氧化所用的 Amplex UltraRed 底物产生强烈的荧光。通过监测相应的荧光强度，该策略可以选择性和灵敏地检测生物硫醇。该基于纳米花的策略还成功地用于可靠地定量人血清中的生物硫醇，显示出其在临床诊断中的巨大潜力。
