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使用钯纳米立方体标记抗体对过敏个体非侵入性样本中的人免疫球蛋白E进行免疫电化学评估。

Immunoelectrochemical assessment of human IgE in non-invasive samples of allergic individuals using PdNCs-labelled antibodies.

作者信息

Rodríguez-Penedo Alejandro, Costa-Rama Estefanía, Pereiro Rosario, Fernández Beatriz, Fernández-Abedul M Teresa

机构信息

Department of Physical and Analytical Chemistry, University of Oviedo, Julian Clavería 8, 33006, Oviedo, Spain.

出版信息

Mikrochim Acta. 2025 Mar 18;192(4):237. doi: 10.1007/s00604-025-07083-3.

Abstract

The escalating global prevalence of allergies presents a substantial public health challenge. Immunoglobulin E (IgE) serves as a key biomarker for allergic diseases, often measured in blood serum by ELISA immunoassays. Despite recent interest in minimally invasive sampling of biological fluids, the low sample volumes and IgE concentrations demand highly sensitive methodologies, typically confined to centralized laboratories. In this article, a decentralizable approach based on competitive immunoassays using Pd nanocluster (PdNCs)-labelled antibodies for electrochemical detection is proposed. With this aim, PdNCs were successfully bioconjugated with an anti-hIgE antibody to perform competitive immunoassays. To improve the analytical capabilities of the methodology, disposable screen-printed carbon electrodes with dual working electrodes were used for enhancing precision. Prior electrodeposition of PdNCs at - 0.6 V for 90 s significantly improved sensitivity (7.1 µA g ng⁻) and lowered the limit of detection (LoD) to 0.3 ng g⁻ for PdNCs determination. The use of PdNCs as labels resulted in an improvement in the LoD for IgE determination. Calibration curves performed using competitive immunoassays for IgE determination, ranging from 10 to 10 ng g, demonstrated sensitivity comparable to high-tech methods, with a LoD of 0.008 ng g for electrochemical measurements. Bimodal detection of Pd (linear sweep voltammetry and inductively coupled plasma-mass spectrometry) in various biological fluids (saliva, tears, nasal exudate, capillary blood, and blood serum) revealed significant differences in IgE levels between allergic and non-allergic individuals. Notably, capillary blood correlated strongly with serum blood, and a certain correlation has also been found with nasal exudate. The electrochemical approach, combining sensitivity and precision with non-invasive sampling, offers a simplified alternative for IgE determination in allergic disease.

摘要

全球过敏患病率不断上升,这对公共卫生构成了重大挑战。免疫球蛋白E(IgE)是过敏性疾病的关键生物标志物,通常通过酶联免疫吸附测定(ELISA)在血清中进行检测。尽管最近人们对生物流体的微创采样很感兴趣,但由于样本量低和IgE浓度低,需要高度灵敏的方法,而这些方法通常局限于集中式实验室。在本文中,我们提出了一种基于竞争性免疫测定的可分散方法,该方法使用钯纳米簇(PdNCs)标记的抗体进行电化学检测。为此,成功地将PdNCs与抗人IgE抗体进行生物偶联,以进行竞争性免疫测定。为了提高该方法的分析能力,使用了带有双工作电极的一次性丝网印刷碳电极来提高精度。在-0.6 V下对PdNCs进行90 s的预电沉积显著提高了灵敏度(7.1 µA g ng⁻),并将PdNCs测定的检测限(LoD)降低至0.3 ng g⁻。使用PdNCs作为标记物提高了IgE测定的LoD。使用竞争性免疫测定法测定IgE的校准曲线范围为10至10 ng g,其灵敏度与高科技方法相当,电化学测量的LoD为0.008 ng g。通过对各种生物流体(唾液、眼泪、鼻分泌物、毛细血管血和血清)中的钯进行双峰检测(线性扫描伏安法和电感耦合等离子体质谱法),发现过敏个体和非过敏个体之间的IgE水平存在显著差异。值得注意的是,毛细血管血与血清血密切相关,并且与鼻分泌物也存在一定的相关性。这种电化学方法将灵敏度和精度与非侵入性采样相结合,为过敏性疾病中IgE的测定提供了一种简化的替代方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0d9/11920355/b81337b7a6f5/604_2025_7083_Fig1_HTML.jpg

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