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Protein Complex Affinity Capture from Cryomilled Mammalian Cells

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Elastomeric negative acoustic contrast particles for affinity capture assays.

Kevin W Cushing¹, Menake E Piyasena, Nick J Carroll

¹Department of Chemical and Nuclear Engineering, Center for Biomedical Engineering, The University of New Mexico, Albuquerque, New Mexico 87131, United States.

Analytical Chemistry

|January 22, 2013

Summary

Researchers developed elastomeric capture microparticles (ECμPs) for acoustophoretic separation. This method enables sensitive detection of biomarkers like prostate specific antigen and immunoglobulin (IgG) using flow cytometry.

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Area of Science:

Biomedical Engineering
Analytical Chemistry
Microfluidics

Background:

Development of novel microparticles for biological assays.
Challenges in separating microparticles from complex biological matrices like blood.
Need for sensitive detection methods for biomarkers.

Purpose of the Study:

To develop elastomeric capture microparticles (ECμPs) for acoustophoretic separation.
To functionalize ECμPs for specific biomarker capture (PSA, IgG).
To demonstrate the utility of ECμPs and acoustophoretic separation in flow cytometry assays.

Main Methods:

Fabrication of ECμPs via cross-linking of silicone precursors.
Surface functionalization of ECμPs with antibodies.
Acoustophoretic separation in a microfluidic device utilizing acoustic contrast differences between ECμPs and blood cells.
Analysis of separated ECμPs using flow cytometry.

Main Results:

ECμPs exhibit negative acoustic contrast, enabling separation from positive contrast blood cells.
Successful functionalization of ECμPs for prostate specific antigen (PSA) and immunoglobulin (IgG) detection.
Achieved nanomolar detection limits for PSA and picomolar detection limits for IgG in various biological samples.

Conclusions:

ECμPs combined with acoustophoretic separation offer an effective method for microparticle-based assays.
This approach facilitates sensitive biomarker detection in complex samples.
Potential for developing rapid, high-throughput diagnostic assays.