Membrane-based microfluidic chip constructed and applied in magnetic microparticle anchoring-and-release digital immunoassay

IF 6.1 1区化学 Q1 CHEMISTRY, ANALYTICAL

Talanta Pub Date : 2025-07-28 DOI:10.1016/j.talanta.2025.128644

Songbai Tian , Tingting Xiang , Xinghu Ji , Fuxiang Zhou , Zhike He

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Abstract

Digital immunoassay is a valuable method for quantifying protein at the single-molecule level. Traditional digital immunoassay (the single-molecule arrays, SiMoAs) involves distributing the antibody-antigen-antibody sandwich immune complex into droplets, but some targets are lost during this process. To address this issue, we developed a magnetic microparticle anchoring-and-release digital immunoassay method for protein detection. We embedded a commercial track-etched polycarbonate (PCTE) membrane into the microfluidic chip for droplet generation, and modified the magnetic microparticles (MMPs) with enzymes for signal production. This method involves two processes: first, the enzymes modified MMPs are anchored in 96-well plates through the target protein. Second, the enzymes modified MMPs were released, and digitally quantified in the membrane-based microfluidic chip. This magnetic microparticle anchoring-and-released digital immunoassay method was successfully applied to the detection of p24 protein. Results showed that p24 could be digitally detected within the range of 1–100 pg/mL with good selectivity and accuracy using this digital method. By reducing signal loss during the digital immunoassay process, this method represents a significant advancement in digital technology.

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膜基微流控芯片的构建及其在磁微粒锚定释放数字免疫分析中的应用

数字免疫分析法是一种在单分子水平上定量蛋白质的有价值的方法。传统的数字免疫测定（单分子阵列，SiMoAs）涉及将抗体-抗原-抗体夹心免疫复合物分布到液滴中，但在此过程中会丢失一些靶标。为了解决这个问题，我们开发了一种用于蛋白质检测的磁性微粒锚定和释放数字免疫分析方法。我们将商业轨迹蚀刻聚碳酸酯（PCTE）膜嵌入微流控芯片中以产生液滴，并用酶修饰磁性微粒子（MMPs）以产生信号。该方法包括两个过程：首先，酶修饰的MMPs通过靶蛋白固定在96孔板中。其次，释放酶修饰的MMPs，并在膜基微流控芯片中进行数字量化。该磁微粒锚定释放数字免疫分析法成功应用于p24蛋白的检测。结果表明，该方法可在1 ~ 100 pg/mL范围内对p24进行数字化检测，具有良好的选择性和准确性。通过减少数字免疫分析过程中的信号损失，该方法代表了数字技术的重大进步。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.