Department of Chemistry, Michigan Technological University , Houghton, Michigan 49931, United States.
Department of Chemistry and Biochemistry, University of Maryland , College Park, Maryland 20892, United States.
ACS Nano. 2017 Feb 28;11(2):2052-2059. doi: 10.1021/acsnano.6b08232. Epub 2017 Feb 3.
Enzyme-based colorimetric assays have been widely used in research laboratories and clinical diagnosis for decades. Nevertheless, as constrained by the performance of enzymes, their detection sensitivity has not been substantially improved in recent years, which inhibits many critical applications such as early detection of cancers. In this work, we demonstrate an enzyme-free signal amplification technique, based on gold vesicles encapsulated with Pd-Ir nanoparticles as peroxidase mimics, for colorimetric assay of disease biomarkers with significantly enhanced sensitivity. This technique overcomes the intrinsic limitations of enzymes, thanks to the superior catalytic efficiency of peroxidase mimics and the efficient loading and release of these mimics. Using human prostate surface antigen as a model biomarker, we demonstrated that the enzyme-free assay could reach a limit of detection at the femtogram/mL level, which is over 10-fold lower than that of conventional enzyme-based assay when the same antibodies and similar procedure were used.
基于酶的比色分析方法在研究实验室和临床诊断中已经被广泛应用了几十年。然而,由于酶的性能受限,近年来其检测灵敏度并没有得到实质性的提高,这限制了许多关键应用,如癌症的早期检测。在这项工作中,我们展示了一种无酶信号放大技术,该技术基于金囊泡封装的 Pd-Ir 纳米颗粒作为过氧化物酶模拟物,可用于比色分析具有显著增强的灵敏度的疾病生物标志物。这种技术克服了酶的固有局限性,这要归功于过氧化物酶模拟物的优越催化效率以及这些模拟物的有效加载和释放。用人前列腺表面抗原作为模型生物标志物,我们证明了无酶测定法可以达到飞克毫升级别的检测限,当使用相同的抗体和类似的方法时,比传统的基于酶的测定法低 10 倍以上。