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Cross-linkage urease nanoparticles: a high-efficiency signal-generation tag for portable pH meter-based electrochemical immunoassay of lipocalin-2 protein diagnostics
Microchimica Acta ( IF 5.3 ) Pub Date : 2020-08-05 , DOI: 10.1007/s00604-020-04466-6
Ai-Li Sun 1 , Qing-An Qi 2 , Li-Juan Zhi 3
Affiliation  

An innovative signal-transduction tag based on cross-linked urease nanoparticles (CLENP) was designed for the development of a pH meter-based immunoassay of lipocalin-2 (LCN2). The CLENP was synthesized with a typical desolvation method using ethanol as desolvation agent, followed by functionalization with polyaspartic acid. The carboxylated CLENP were used as the signal-generation tags for the labelling of secondary antibodies via the carbodiimide coupling. Upon target LCN2 introduction, a sandwich-type immune reaction was performed between capture antibody-coated plate and the labeled secondary antibody on the CLENP. The conjugated CLENP in the microplate hydrolyzed urea into ammonia (NH4+) and carbonate (CO32−), resulting in the pH change of solution, which was determined with a handheld pH meter. The pH variation was proportional to target concentration in the sample. By monitoring the pH variation of the urea solution, the level of LCN2 at a concentration as low as 5.2 pg mL−1 was evaluated. The pH meter-based electrochemical immunoassay can be utilized for mass production of miniaturized lab-on-a-chip devices with handheld pH meter, thereby opening new opportunities for protein diagnostics and biosecurity. Graphical abstract An innovative signal-transduction tag based on cross-linked urease nanoparticles was designed for high-efficiency immunoassay of lipocalin-2 with pH meter readout. An innovative signal-transduction tag based on cross-linked urease nanoparticles was designed for high-efficiency immunoassay of lipocalin-2 with pH meter readout.

中文翻译:

交联脲酶纳米颗粒:一种高效信号生成标签,用于基于便携式 pH 计的 lipocalin-2 蛋白电化学免疫分析诊断

基于交联脲酶纳米颗粒 (CLENP) 的创新信号转导标签设计用于开发基于 pH 计的 lipocalin-2 (LCN2) 免疫分析。CLENP 是用典型的去溶剂化方法合成的,使用乙醇作为去溶剂化剂,然后用聚天冬氨酸进行功能化。羧化 CLENP 被用作信号生成标签,用于通过碳二亚胺偶联标记二抗。在引入目标 LCN2 后,在捕获抗体包被的板和 CLENP 上标记的二抗之间进行夹心型免疫反应。微孔板中的共轭 CLENP 将尿素水解成氨 (NH4+) 和碳酸盐 (CO32-),导致溶液的 pH 值发生变化,这可以用手持式 pH 计测定。pH 变化与样品中的目标浓度成正比。通过监测尿素溶液的 pH 值变化,评估了低至 5.2 pg mL-1 的 LCN2 水平。基于 pH 计的电化学免疫测定可用于大规模生产带有手持式 pH 计的微型芯片实验室设备,从而为蛋白质诊断和生物安全开辟新的机遇。图形摘要 基于交联脲酶纳米颗粒的创新信号转导标签被设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。基于交联脲酶纳米颗粒的创新信号转导标签设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。评估了低至 5.2 pg mL-1 的 LCN2 水平。基于 pH 计的电化学免疫测定可用于大规模生产带有手持式 pH 计的微型芯片实验室设备,从而为蛋白质诊断和生物安全开辟新的机遇。图形摘要 基于交联脲酶纳米颗粒的创新信号转导标签被设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。基于交联脲酶纳米颗粒的创新信号转导标签设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。评估了低至 5.2 pg mL-1 的 LCN2 水平。基于 pH 计的电化学免疫测定可用于大规模生产带有手持式 pH 计的微型芯片实验室设备,从而为蛋白质诊断和生物安全开辟新的机遇。图形摘要 基于交联脲酶纳米颗粒的创新信号转导标签被设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。基于交联脲酶纳米颗粒的创新信号转导标签设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。从而为蛋白质诊断和生物安全开辟了新的机会。图形摘要 基于交联脲酶纳米颗粒的创新信号转导标签被设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。基于交联脲酶纳米颗粒的创新信号转导标签设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。从而为蛋白质诊断和生物安全开辟了新的机会。图形摘要 基于交联脲酶纳米颗粒的创新信号转导标签被设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。基于交联脲酶纳米颗粒的创新信号转导标签设计用于具有 pH 计读数的 lipocalin-2 的高效免疫测定。
更新日期:2020-08-05
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