Key Laboratory of Analysis and Detection for Food Safety (Fujian Province & Ministry of Education), Institute of Nanomedicine and Nanobiosensing, Department of Chemistry, Fuzhou University, Fuzhou 350116, PR China.
Institute of Environmental and Analytical Science, School of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004, Henan, PR China.
Biosens Bioelectron. 2015 Aug 15;70:194-201. doi: 10.1016/j.bios.2015.03.039. Epub 2015 Mar 17.
The development of signal-amplified colorimetric immunoassay relies on the design of highly efficient signal-transduction tags. One promising route is to exploit a novel enzyme mimetic system as the signal label. Herein, we report that urchin-like (gold core)@(platinum shell) nanohybrids (Au@PtNHs) can be utilized as a highly efficient peroxidase mimetic system for in situ amplified colorimetric immunoassay of prostate-specific antigen (PSA, one kind of tumor marker). Initially, urchin-like Au@PtNHs were discovered to outperform horseradish peroxidase (HRP) by a vast margin in terms of the turnover number toward hydrogen peroxide (H2O2)-3,3',5,5'-tetramethylbenzidine (TMB) system and the stability against high temperatures and HRP inhibitors. Based on this discovery, the assay was simply carried out on a capture antibody-immobilized microplate by using the Au@PtNH-labeled detection antibody as a signal-transduction tag with a sandwich-type assay mode. The colorimetric signal stemmed from the labeled Au@PtNHs toward catalytic oxidation of TMB-H2O2 system. Experimental results indicated that the Au@PtNH-based colorimetric immunoassay could display a good colorimetric response toward PSA in the dynamic working range of 5-500 pg mL(-1) with a low detection limit of 2.9 pg mL(-1). Meanwhile, the developed immunoassay exhibited good precision and reproducibility, high specificity and acceptable accuracy for the detection of clinical serum samples. These results open up a new horizon for the development of highly sensitive, highly stable and inexpensive non-enzyme immunoassay platforms as an alternative to conventional enzyme-based immunoassay platforms.
基于信号放大的比色免疫分析的发展依赖于高效信号转导标签的设计。一种很有前途的方法是利用一种新型的酶模拟体系作为信号标记。本文报道了,具有仿贻贝结构的(金核)@(铂壳)纳米杂化材料(Au@PtNHs)可用作高效过氧化物酶模拟体系,用于前列腺特异性抗原(PSA,一种肿瘤标志物)的原位放大比色免疫分析。首先,与辣根过氧化物酶(HRP)相比,具有仿贻贝结构的 Au@PtNHs 在过氧化氢(H2O2)-3,3',5,5'-四甲基联苯胺(TMB)体系中的周转数和对高温及 HRP 抑制剂的稳定性方面具有显著优势。基于这一发现,该测定方法简单地在捕获抗体固定的微孔板上进行,采用 Au@PtNH 标记的检测抗体作为信号转导标签,以三明治型测定模式进行。比色信号源于标记的 Au@PtNHs 对 TMB-H2O2 体系的催化氧化。实验结果表明,基于 Au@PtNH 的比色免疫分析对 PSA 具有良好的比色响应,动态工作范围为 5-500 pg mL(-1),检测限低至 2.9 pg mL(-1)。同时,所开发的免疫分析方法具有良好的精密度和重现性、高特异性和可接受的临床血清样本检测准确性。这些结果为开发高灵敏度、高稳定性和低成本的非酶免疫分析平台开辟了新的前景,可作为传统酶免疫分析平台的替代方案。