Elastomeric Negative Acoustic Contrast Particles for Affinity Capture Assays

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• 作者：Kevin W. Cushing ; Menake E. Piyasena ; Nick J. Carroll ; Gian C. Maestas ; Beth Ann L贸pez ; Bruce S. Edwards ; Steven W. Graves ; Gabriel P. L贸pez
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：February 19, 2013
• 年：2013
• 卷：85
• 期：4
• 页码：2208-2215
• 全文大小：351K
• 年卷期：v.85,no.4(February 19, 2013)
• ISSN：1520-6882

文摘

This report describes the development of elastomeric capture microparticles (EC渭Ps) and their use with acoustophoretic separation to perform microparticle assays via flow cytometry.We have developed simple methods to form EC渭Ps by cross-linking droplets of common commercially available silicone precursors in suspension followed by surface functionalization with biomolecular recognition reagents. The EC渭Ps 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 EC渭Ps 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 EC渭Ps at the antinodes. Laminar flow of the separated particles to downstream collection ports allows for collection of the separated negative contrast (EC渭Ps) and positive contrast particles (cells). Separated EC渭Ps 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.