Antibody Surface Coverage Drives Matrix Interference in Microfluidic Capillary Immunoassays.
Antibody Surface Coverage Drives Matrix Interference in Microfluidic Capillary Immunoassays.
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DOI：
10.1021/acssensors.1c00704
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发表时间：
2021-07-23
期刊：
影响因子：
8.9
通讯作者：
Reis NM
中科院分区：
文献类型：
--
作者：
Barbosa AI;Edwards AD;Reis NM
生物传感器的性能通常在缓冲液中进行优化，这在生物样品的应用期间带来不一致性。用于最小化样品（基质）干扰的当前策略对于自动化和简化是复杂的，涉及例如，样品稀释或血清/血浆回收。这项研究首次使用数百个实际的微流控免疫测定含氟聚合物条进行系统分析，以了解微流系统中的基质干扰。由于许多干扰因子具有检测特异性，因此我们探索了一系列酶免疫检测的基质干扰，包括直接mIgG/抗mIgG、夹心癌症生物标志物PSA和夹心炎性细胞因子IL-1β。血清基质干扰显著受毛细管抗体表面覆盖的影响，首次表明血清基质效应的主要原因是低亲和力血清组分（例如，自身抗体）与高亲和力抗原竞争固定化抗体。使用不同毛细管直径进行的其他实验证实了抗体表面覆盖在管理基质干扰中的重要性。基于这些发现，我们提出了一种新的分析方法，其中抗体表面覆盖和样品孵育时间是消除和/或最小化血清基质干扰的关键，包括在血清而不是缓冲液中进行的生物测定优化，而不会影响生物测定的性能或增加额外的成本或步骤。这将有助于建立一条新的途径，以更快地开发现代即时检测和有效的生物传感器。
The performance of biosensors is often optimized in buffers, which brings inconsistencies during applications with biological samples. Current strategies for minimizing sample (matrix) interference are complex to automate and miniaturize, involving, e.g., sample dilution or recovery of serum/plasma. This study shows the first systematic analysis using hundreds of actual microfluidic immunoassay fluoropolymer strips to understand matrix interference in microflow systems. As many interfering factors are assay-specific, we have explored matrix interference for a range of enzymatic immunoassays, including a direct mIgG/anti-mIgG, a sandwich cancer biomarker PSA, and a sandwich inflammatory cytokine IL-1β. Serum matrix interference was significantly affected by capillary antibody surface coverage, suggesting for the first time that the main cause of the serum matrix effect is low-affinity serum components (e.g., autoantibodies) competing with high-affinity antigens for the immobilized antibody. Additional experiments carried out with different capillary diameters confirmed the importance of antibody surface coverage in managing matrix interference. Building on these findings, we propose a novel analytical approach where antibody surface coverage and sample incubation times are key for eliminating and/or minimizing serum matrix interference, consisting in bioassay optimization carried out in serum instead of buffer, without compromising the performance of the bioassay or adding extra cost or steps. This will help establishing a new route toward faster development of modern point-of-care tests and effective biosensor development.
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