Development of Simple and Rapid Bead-Based Cytometric Immunoassays Using Superparamagnetic Hybrid Core–Shell Microparticles

IF 4.6 Q1 CHEMISTRY, ANALYTICAL
Charlie Tobias, Daniel López-Puertollano, Antonio Abad-Somovilla, Josep V. Mercader, Antonio Abad-Fuentes, Knut Rurack
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引用次数: 0

Abstract

Flow cytometry-based immunoassays are valuable in biomedical research and clinical applications due to their high throughput and multianalyte capability, but their adoption in areas such as food safety and environmental monitoring is limited by long assay times and complex workflows. Rapid, simplified bead-based cytometric immunoassays are needed to make these methods viable for point-of-need applications, especially with the increasing accessibility of miniaturized cytometers. This work introduces superparamagnetic hybrid polystyrene-silica core–shell microparticles as promising alternatives to conventional polymer beads in competitive cytometric immunoassays. These beads, featuring high specificity, sensitivity, and excellent handling capabilities via magnetic separation, were evaluated with three different antibodies and binding methods, showing variations in signal intensity based on the antibody and its attachment method. The optimal performance was achieved through a secondary antibody binding approach, providing strong and consistent signals with minimal uncertainty. The optimized protocol made it possible to achieve a detection limit of 0.025 nM in a total assay time of only 15 min and was successfully used to detect ochratoxin A (OTA) in raw flour samples. This work highlights the potential of these beads as versatile tools for flow cytometry-based immunoassays, with significant implications for food safety, animal health, environmental monitoring, and clinical diagnostics.

Abstract Image

利用超顺磁性混合核壳微粒开发简单快速的珠基细胞免疫测定方法
基于流式细胞仪的免疫测定因其高通量和多分析能力而在生物医学研究和临床应用中具有重要价值,但其在食品安全和环境监测等领域的应用却受到化验时间长和工作流程复杂的限制。需要快速、简化的基于微珠的细胞计量免疫测定,使这些方法在需求点应用中切实可行,特别是随着微型细胞仪的日益普及。这项工作介绍了超顺磁性混合聚苯乙烯-二氧化硅核壳微粒,作为竞争性细胞免疫测定中传统聚合物微珠的有前途的替代品。这些珠子具有高特异性、高灵敏度和通过磁性分离的出色处理能力,我们用三种不同的抗体和结合方法对其进行了评估,结果表明,根据抗体及其附着方法的不同,信号强度也有差异。通过二抗结合的方法实现了最佳性能,可提供强而稳定的信号,且不确定性最小。优化后的方案使检测限达到了 0.025 nM,总检测时间仅为 15 分钟,并成功用于检测生面粉样品中的赭曲霉毒素 A(OTA)。这项工作凸显了这些微珠作为流式细胞仪免疫测定多功能工具的潜力,对食品安全、动物健康、环境监测和临床诊断具有重要意义。
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来源期刊
ACS Measurement Science Au
ACS Measurement Science Au 化学计量学-
CiteScore
5.20
自引率
0.00%
发文量
0
期刊介绍: ACS Measurement Science Au is an open access journal that publishes experimental computational or theoretical research in all areas of chemical measurement science. Short letters comprehensive articles reviews and perspectives are welcome on topics that report on any phase of analytical operations including sampling measurement and data analysis. This includes:Chemical Reactions and SelectivityChemometrics and Data ProcessingElectrochemistryElemental and Molecular CharacterizationImagingInstrumentationMass SpectrometryMicroscale and Nanoscale systemsOmics (Genomics Proteomics Metabonomics Metabolomics and Bioinformatics)Sensors and Sensing (Biosensors Chemical Sensors Gas Sensors Intracellular Sensors Single-Molecule Sensors Cell Chips Arrays Microfluidic Devices)SeparationsSpectroscopySurface analysisPapers dealing with established methods need to offer a significantly improved original application of the method.
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