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A microfluidic electrochemiluminescent device for detecting cancer biomarker proteins

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Abstract

We describe an electrochemiluminescence (ECL) immunoarray incorporated into a prototype microfluidic device for highly sensitive protein detection and apply this system to accurate, sensitive measurements of prostate-specific antigen (PSA) and interleukin-6 (IL-6) in serum. The microfluidic system employed three molded polydimethylsiloxane (PDMS) channels on a conductive pyrolytic graphite chip (2.5 × 2.5 cm) inserted into a machined chamber and interfaced with a pump, switching valve, and sample injector. Each of the three PDMS channels encompasses three 3 μL analytical wells. Capture-antibody-decorated single-wall carbon nanotube forests are fabricated in the bottom of the wells. The antigen is captured by these antibodies on the well bottoms. Then, a RuBPY-silica-secondary antibody (Ab2) label is injected to bind to antigen on the array, followed by injection of sacrificial reductant tripropylamine (TPrA) to produce ECL. For detection, the chip is placed into an open-top ECL measuring cell, and the channels are in contact with electrolyte in the chamber. Potential applied at 0.95 V versus Ag/AgCl oxidizes TPrA to produce ECL by redox cycling the RuBPY species in the particles, and ECL light is measured by a charge-coupled device camera. This approach achieved ultralow detection limits of 100 fg mL−1 for PSA (9 zeptomole) and 10 fg mL−1 (1 zeptomole) for IL-6 in calf serum, a 10–25-fold improvement of a similar non-microfluidic array. PSA and IL-6 in synthetic cancer patient serum samples were detected in 1.1 h and results correlated well with single-protein enzyme-linked immunosorbent assays.

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Acknowledgments

This work was supported by grant no. EB014586 from the National Institute of Biomedical Imaging and Bioengineering, NIH. We also acknowledge FAPESP Proc. No. 11/02259-6 for financial support to Dr. R.C. Faria for a sabbatical visit.

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Authors and Affiliations

  1. Department of Chemistry, University of Connecticut, Storrs, CT, 06269, USA

    Naimish P. Sardesai, Karteek Kadimisetty, Ronaldo Faria & James F. Rusling

  2. Departamento de Química, Universidade Federal de São Carlos, São Carlos, SP, Brazil

    Ronaldo Faria

  3. Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, 06032, USA

    James F. Rusling

  4. Institute of Materials Science, University of Connecticut, Storrs, CT, 06269, USA

    James F. Rusling

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  1. Naimish P. Sardesai
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Correspondence to James F. Rusling.

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Published in the topical collection Bioelectroanalysis with guest editors Nicolas Plumeré, Magdalena Gebala, and Wolfgang Schuhmann.

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Sardesai, N.P., Kadimisetty, K., Faria, R. et al. A microfluidic electrochemiluminescent device for detecting cancer biomarker proteins. Anal Bioanal Chem 405, 3831–3838 (2013). https://doi.org/10.1007/s00216-012-6656-5

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  • DOI: https://doi.org/10.1007/s00216-012-6656-5

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