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Elastomeric Negative Acoustic Contrast Particles for Affinity Capture Assays

机译:弹性负声学造影颗粒的亲和捕获试验

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摘要

This report describes the development of elastomeric capture microparticles (ECμPs) and their use with acoustophoretic separation to perform microparticle assays via flow cytometry. We have developed simple methods to form ECμPsby crosslinking droplets of common commercially available silicone precursors in suspension followed by surface functionalization with biomolecular recognition reagents. The ECμPs are compressible particles that exhibit negative acoustic contrast in ultrasound when suspended in aqueous media, blood serum or diluted blood. In this study, these particles have been functionalized with antibodies to bind prostate specific antigen and immunoglobulin (IgG). Specific separation of the ECμPs from blood cells is achieved by flowing them through a microfluidic acoustophoretic device that uses an ultrasonic standing wave to align the blood cells, which exhibit positive acoustic contrast, at a node in the acoustic pressure distribution while aligning the negative acoustic contrast ECμPs at the antinodes. Laminar flow of the separated particles to downstream collection ports allows for collection of the separated negative contrast (ECμPs) and positive contrast particles (cells). Separated ECμPs were analyzed via flow cytometry to demonstrate nanomolar detection for prostate specific antigen in aqueous buffer and picomolar detection for IgG in plasma and diluted blood samples. This approach has potential applications in the development of rapid assays that detect the presence of low concentrations of biomarkers in a number of biological sample types.
机译:这份报告描述了弹性捕获微粒(ECμP)的发展及其在声电泳分离中通过流式细胞仪进行微粒测定的用途。我们已经开发出一种简单的方法,可以通过将悬浮的通用市售有机硅前体的液滴交联,然后用生物分子识别试剂进行表面官能化来形成ECμPs。 ECμPs是可压缩颗粒,当悬浮在水性介质,血清或稀释的血液中时,在超声中显示出负面的声学对比。在这项研究中,这些颗粒已通过抗体功能化以结合前列腺特异性抗原和免疫球蛋白(IgG)。 ECμPs与血细胞的特定分离是通过使它们流过微流声电泳设备实现的,该设备使用超声波驻波将声压分布呈正值的血细胞排列在声压分布的节点上,同时排列负的声对比度,从而显示出正的声对比度。波腹处的ECμP。分离的颗粒向下游收集口的层流允许收集分离的负对比剂(ECμPs)和正对比剂颗粒(细胞)。通过流式细胞仪分析分离的ECμP,以证明在水溶液缓冲液中检测纳摩尔浓度的前列腺特异性抗原,并在血浆和稀释的血液样本中检测皮摩尔的IgG。这种方法在快速检测方法的开发中具有潜在的应用价值,该方法可以检测多种生物样品类型中低浓度的生物标志物的存在。

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