A.N. Bach Institute of Biochemistry, Federal Research Centre "Fundamentals of Biotechnology", Russian Academy of Sciences, Leninsky Prospect 33, 119071, Moscow, Russia.
Mikrochim Acta. 2021 Aug 27;188(9):309. doi: 10.1007/s00604-021-04968-x.
Platinum-containing nanozymes with peroxidase-mimicking activity (PMA) have found a broad application in bioanalytical methods and are potentially able to compete with enzymes as the labels. However, traditionally used methods for the synthesis of nanozymes result in only a small fraction of surface-exposed Pt atoms, which participate in catalysis. To overcome this limitation, we propose a new approach for the synthesis of nanozymes with the efficient dispersion of Pt atoms on particles' surfaces. The synthesis of nanozymes includes three steps: the synthesis of gold nanoparticles (Au NPs), the overgrowth of a silver layer over Au NPs (Au@Ag NPs, 6 types of NPs with different thicknesses of Ag shell), and the galvanic replacement of silver with PtCl leading to the formation of trimetallic Au@Ag-Pt NPs with uniformly deposited catalytic sites and high Pt-utilization efficiency. Au@Ag-Pt NPs (23 types of NPs with different concentrations of Pt) with various sizes, morphology, optical properties, and PMA were synthesized and comparatively tested. Using energy-dispersive spectroscopy mapping, we confirm the formation of core@shell Au@Ag NPs and dispersion of surface-exposed Pt. The selected Au@Ag-Pt NPs were conjugated with monoclonal antibodies and used as the colorimetric and catalytic labels in lateral flow immunoassay of the inflammation biomarker: C-reactive protein (CRP). The colorimetric signal enhancement was achieved by the oxidation of 3,3'-diaminobenzidine by HO catalyzed by Au@Ag-Pt NPs directly on the test strip. The use of Au@Ag-Pt NPs as the catalytic label produces a 65-fold lower limit of CRP detection in serum (15 pg mL) compared with Au NPs and ensures the lowest limit of detection for equipment-free lateral flow immunoassays. The assay shows a high correlation with data of enzyme-linked immunosorbent assay (R = 0.986) and high recovery (83.7-116.2%) in serum and plasma. The assay retains all the benefits of lateral flow immunoassay as a point-of-care method.
具有过氧化物酶模拟活性 (PMA) 的含铂纳米酶在生物分析方法中得到了广泛的应用,并且有可能与酶一起作为标记物使用。然而,传统的纳米酶合成方法只能得到一小部分参与催化的表面暴露的 Pt 原子。为了克服这一限制,我们提出了一种新的方法来合成具有高效分散 Pt 原子在颗粒表面的纳米酶。纳米酶的合成包括三个步骤:金纳米粒子 (Au NPs) 的合成、Au NPs 上银层的过度生长 (Au@Ag NPs,具有 6 种不同厚度银壳的 NPs) 以及银与 PtCl 的电置换,导致形成具有均匀沉积催化位点和高 Pt 利用率的三元 Au@Ag-Pt NPs。合成了具有不同尺寸、形态、光学性质和 PMA 的 23 种不同 Pt 浓度的 Au@Ag-Pt NPs,并进行了比较测试。通过能谱映射,我们证实了核壳 Au@Ag NPs 的形成和表面暴露的 Pt 的分散。选择的 Au@Ag-Pt NPs 与单克隆抗体偶联,并用作炎症生物标志物 C-反应蛋白 (CRP) 的侧向流动免疫分析的比色和催化标记物。通过 Au@Ag-Pt NPs 直接在测试条上催化 HO 氧化 3,3'-二氨基联苯胺实现了比色信号的增强。与 Au NPs 相比,Au@Ag-Pt NPs 作为催化标记物可将 CRP 在血清中的检测下限降低 65 倍(15 pg mL),并确保无设备侧向流动免疫分析的最低检测限。该测定法与酶联免疫吸附测定法的数据具有高度相关性 (R = 0.986),并且在血清和血浆中的回收率高 (83.7-116.2%)。该测定法保留了侧向流动免疫分析作为即时检测方法的所有优势。
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