{"title":"使用pdncs标记抗体对非侵入性过敏个体样本中的人IgE进行免疫电化学评价","authors":"Alejandro Rodríguez-Penedo, Estefanía Costa-Rama, Rosario Pereiro, Beatriz Fernández, M. Teresa Fernández-Abedul","doi":"10.1007/s00604-025-07083-3","DOIUrl":null,"url":null,"abstract":"<div><p>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⁻<sup>1</sup>) and lowered the limit of detection (LoD) to 0.3 ng g⁻<sup>1</sup> 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<sup>−5</sup> to 10<sup>2</sup> ng g<sup>−1</sup>, demonstrated sensitivity comparable to high-tech methods, with a LoD of 0.008 ng g<sup>−1</sup> 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.</p><h3>Graphical Abstract</h3>\n<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":705,"journal":{"name":"Microchimica Acta","volume":"192 4","pages":""},"PeriodicalIF":5.3000,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s00604-025-07083-3.pdf","citationCount":"0","resultStr":"{\"title\":\"Immunoelectrochemical assessment of human IgE in non-invasive samples of allergic individuals using PdNCs-labelled antibodies\",\"authors\":\"Alejandro Rodríguez-Penedo, Estefanía Costa-Rama, Rosario Pereiro, Beatriz Fernández, M. Teresa Fernández-Abedul\",\"doi\":\"10.1007/s00604-025-07083-3\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>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⁻<sup>1</sup>) and lowered the limit of detection (LoD) to 0.3 ng g⁻<sup>1</sup> 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<sup>−5</sup> to 10<sup>2</sup> ng g<sup>−1</sup>, demonstrated sensitivity comparable to high-tech methods, with a LoD of 0.008 ng g<sup>−1</sup> 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. 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引用次数: 0
摘要
全球过敏症发病率的不断上升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ELISA免疫分析在血清中测量。尽管最近对生物液体的微创取样很感兴趣,但低样本量和IgE浓度需要高度敏感的方法,通常仅限于集中实验室。本文提出了一种基于竞争性免疫分析的分散方法,使用Pd纳米簇(pdnc)标记的抗体进行电化学检测。为此,我们成功地将pdcs与抗hige抗体进行了生物偶联,以进行竞争性免疫测定。为了提高该方法的分析能力,采用了一次性丝网印刷碳电极双工作电极来提高精度。预先在- 0.6 V下电沉积pddc 90 s,可以显著提高灵敏度(7.1µA g⁻1),并将pddc的检测限(LoD)降低到0.3 ng g⁻1。使用PdNCs作为标记可改善IgE测定的LoD。使用竞争性免疫分析法进行IgE测定的校准曲线,范围从10−5到102 ng g−1,显示出与高科技方法相当的灵敏度,电化学测量的LoD为0.008 ng g−1。对各种生物体液(唾液、眼泪、鼻渗出液、毛细血管血和血清)进行双模态Pd检测(线性扫描伏安法和电感耦合血浆质谱法),发现过敏和非过敏个体之间IgE水平存在显著差异。值得注意的是,毛细血管血与血清血有很强的相关性,与鼻渗出液也有一定的相关性。电化学方法,结合灵敏度和精度与非侵入性采样,提供了一种简单的替代IgE检测过敏性疾病。图形抽象
Immunoelectrochemical assessment of human IgE in non-invasive samples of allergic individuals using PdNCs-labelled antibodies
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⁻1) and lowered the limit of detection (LoD) to 0.3 ng g⁻1 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 10−5 to 102 ng g−1, demonstrated sensitivity comparable to high-tech methods, with a LoD of 0.008 ng g−1 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.
期刊介绍:
As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.