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基于过氧化物酶模拟纳米酶免疫测定的高敏比色法检测过敏。

Highly sensitive colorimetric detection of allergies based on an immunoassay using peroxidase-mimicking nanozymes.

机构信息

Department of BioNano Technology, Gachon University, Seongnam, Gyeonggi 13120, Republic of Korea.

出版信息

Analyst. 2018 Feb 26;143(5):1182-1187. doi: 10.1039/c7an01866e.

DOI:10.1039/c7an01866e
PMID:29405213
Abstract

Nanomaterials that exhibit enzyme-like characteristics, which are called nanozymes, have recently attracted significant attention due to their potential to overcome the intrinsic limitations of natural enzymes, such as low stability and relatively high cost for preparation and purification. In this study, we report a highly efficient colorimetric allergy detection system based on an immunoassay utilizing the peroxidase-mimicking activity of hierarchically structured platinum nanoparticles (H-Pt NPs). The H-Pt NPs had a diameter of 30 nm, and were synthesized by a seed-mediated growth method, which led to a significant amount of peroxidase-like activity. This activity mainly occurs because of the high catalytic power of the Pt element, and the fact that the H-Pt NPs have a large surface area available for catalytic events. The H-Pt NPs were conjugated to an antibody for the detection of immunoglobulin E (IgE) in the analytes; IgE is a representative marker for the diagnosis of allergies. They were then successfully integrated into a conventionally used allergy diagnostic test, the ImmunoCAP diagnostic test, as a replacement for natural signaling enzymes. Using this strategy, total and specific IgE levels were detected within 5 min at room temperature, with high specificity and sensitivity. The practical utility of the immunoassay was also successfully verified by correctly determining the levels of both total and specific IgE in real human serum samples with high precision and reproducibility. The present H-Pt NP-based immunoassay system would serve as a platform for rapid, robust, and convenient analysis of IgE, and can be extended to the construction of diagnostic systems for a variety of clinically important target molecules.

摘要

纳米材料具有酶的特性,被称为纳米酶,由于其克服天然酶固有局限性的潜力,如稳定性低和制备及纯化成本相对较高,最近引起了广泛关注。在本研究中,我们报告了一种基于免疫测定的高效比色过敏检测系统,该系统利用分级结构铂纳米粒子(H-Pt NPs)的过氧化物酶模拟活性。H-Pt NPs 的直径为 30nm,通过种子介导的生长方法合成,具有大量过氧化物酶样活性。这种活性主要是由于 Pt 元素的高催化能力,以及 H-Pt NPs 具有大量可用于催化事件的表面积。H-Pt NPs 与抗体偶联,用于检测分析物中的免疫球蛋白 E(IgE);IgE 是过敏诊断的代表性标志物。然后,它们成功地整合到常规使用的过敏诊断测试——ImmunoCAP 诊断测试中,作为天然信号酶的替代品。使用这种策略,在室温下 5 分钟内即可检测总 IgE 和特异性 IgE,具有高特异性和灵敏度。通过高精度和可重复性,该免疫测定在正确确定真实人血清样本中的总 IgE 和特异性 IgE 水平方面也成功验证了其实用性。本研究基于 H-Pt NP 的免疫分析系统将成为快速、稳健、方便的 IgE 分析平台,并可扩展到构建各种临床重要靶分子的诊断系统。

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