Exploitation of Luminescent Lanthanide Nanoparticles for a Sensitivity-Enhanced ELISA Detection Method,Analytical Chemistry

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Exploitation of Luminescent Lanthanide Nanoparticles for a Sensitivity-Enhanced ELISA Detection Method
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-26 , DOI: 10.1021/acs.analchem.3c04821
Ali A Kassir ₁ , Clémence Cheignon ₁ , Loïc J Charbonnière ₁

Affiliation

A new detection method based on the photoluminescence properties of dye-sensitized lanthanide nanoparticles (Ln NPs) was developed for enzyme-linked immunosorbent assays (ELISAs). In this method, the horseradish peroxidase (HRP) enzyme catalyzes the oxidation of phenol derivatives in the presence of hydrogen peroxide, providing dimers that are able to interact with the Ln NP surface and to efficiently photosensitize the Ln ions. Due to the very long emission lifetime of Ln, the time-gated detection of Ln NP luminescence allows the elimination of background noise due to the biological environment. After a comparison of the enzyme-catalyzed oxidation of various phenol derivatives, methyl 4-hydroxyphenyl acetate (MHPA) was selected as the most promising substrate, as the highest Ln emission intensity was observed following its HRP-catalyzed oxidation. After a meticulous optimization of the conditions of both the enzymatic reaction and the Ln sensitization (buffer, pH, concentration of the reactants, NP type, etc.), this new detection method was successfully implemented in a commercial insulin ELISA kit as a proof-of-concept, with an increased sensitivity compared to the commercial detection method.

中文翻译：

利用发光稀土纳米颗粒提高灵敏度 ELISA 检测方法

开发了一种基于染料敏化稀土纳米颗粒（Ln NP）光致发光特性的新检测方法，用于酶联免疫吸附测定（ELISA）。在该方法中，辣根过氧化物酶 (HRP) 在过氧化氢存在下催化苯酚衍生物的氧化，提供能够与 Ln NP 表面相互作用并有效光敏化 Ln 离子的二聚体。由于 Ln 的发射寿命非常长，Ln NP 发光的时间选通检测可以消除生物环境引起的背景噪声。在对各种苯酚衍生物的酶催化氧化进行比较后，选择 4-羟基苯乙酸甲酯 (MHPA) 作为最有前途的底物，因为在其 HRP 催化氧化后观察到最高的 Ln 发射强度。经过对酶反应和 Ln 敏化条件（缓冲液、pH、反应物浓度、NP 类型等）的精心优化，这种新的检测方法成功应用于商业胰岛素 ELISA 试剂盒，作为证明-与商业检测方法相比，其灵敏度更高。

更新日期：2024-01-26

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