Elastomeric Negative Acoustic Contrast Particles for Affinity Capture Assays

Elastomeric Negative Acoustic Contrast Particles for Affinity Capture Assays

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 ECμPs by cross-linking droplets of common commercially available silicone prec...

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Bibliographic Details
Published in:Analytical chemistry (Washington) Vol. 85; no. 4; pp. 2208 - 2215
Main Authors: Cushing, Kevin W, Piyasena, Menake E, Carroll, Nick J, Maestas, Gian C, López, Beth Ann, Edwards, Bruce S, Graves, Steven W, López, Gabriel P
Format: Journal Article
Language:English
Published: United States American Chemical Society 19-02-2013
Subjects:
Animals
Antibodies, Monoclonal - immunology
Antigens
Aqueous solutions
Biomarkers - analysis
Biomarkers - blood
Cells
Dimethylpolysiloxanes - chemistry
Elastomers
Flow cytometry
Flow Cytometry - methods
Humans
Immunoglobulin G - blood
Immunoglobulins
Mice
Microfluidic Analytical Techniques
Microspheres
Polymers - chemical synthesis
Polymers - chemistry
Prostate
Prostate-Specific Antigen - analysis
Swine
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Summary: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 ECμPs by cross-linking 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.
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Current Address: Harvard School of Engineering and Applied Sciences, Pierce Hall, 29 Oxford Street, Cambridge, MA 02138, USA.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac3029344