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Isolation and detection of single molecules on paramagnetic beads using sequential fluid flows in microfabricated

Cheuk W Kan¹, Andrew J Rivnak, Todd G Campbell

¹Quanterix Corporation, Cambridge, MA 02139, USA.

|December 20, 2011

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This study presents a novel method for isolating single enzyme-labeled proteins on paramagnetic beads within femtolitre wells. This digital ELISA technique enables sensitive biomarker detection for medical diagnosis.

Area of Science:

Biotechnology
Analytical Chemistry
Materials Science

Background:

Developing sensitive protein detection methods is crucial for biomarker validation and medical diagnosis.
Existing digital ELISA methods often face limitations in manufacturing, automation, and cost-effectiveness.

Purpose of the Study:

To report a new method for isolating individual paramagnetic beads in femtolitre wells for single-molecule protein detection.
To evaluate the analytical performance of this novel device for digital ELISA applications.

Main Methods:

Fabrication of femtolitre well arrays in cyclic olefin polymer (COP) using injection molding.
Sequential fluidic flows for bead loading, excess bead removal, and well isolation using fluorocarbon oil.
Detection of single enzyme-labeled proteins via fluorescence imaging and digital ELISA.

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Main Results:

Successful isolation of single paramagnetic beads in femtolitre wells.
Demonstrated sensitive detection of prostate specific antigen (PSA) from 0.011 pg mL(-1) to 100 pg mL(-1).
Analytical performance favorably compared to standard glass array methods.

Conclusions:

The enclosed fluidic device offers advantages for low-cost manufacturing and automation.
This technology has potential for biomarker validation and medical diagnosis.
Enables development of new instruments for sensitive protein analysis.