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Development of an ultrafast quantitative heterogeneous immunoassay on pre-functionalized poly(dimethylsiloxane) microfluidic chips for the next-generation immunosensors

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Abstract

Although the reaction time for antigen-antibody binding has been greatly reduced in microchannels, other processes in heterogeneous immunoassays (HEIs), such as blocking and antigen adsorption have not benefited from miniaturization as a reduction in size to micro dimensions does not increase the speed of these processes significantly. The overall assay time of reported microfluidic HEIs has continued to be limited by these processes. In this study, we successfully develop an ultrafast quantitative HEI with pre-functionalized microfluidic poly(dimethylsiloxane) (PDMS) chips. The protein A functionalized PDMS surface is found to be highly effective in reducing the antigen adsorption time in microchannels. The functionalized surfaces can be stable at least for 2.5 months when stored at 4°C in a buffer solution consisting of 10 mM Tris, 0.05% bovine serum albumin, 0.05% Proclin 300, and 5% glycerol. In addition, the immunosorption process, which is substantially accelerated in micro scale, results in a significant reduction in nonspecific binding. The time of blocking step can therefore be reduced to a minimum or can be eliminated. The overall assay for detecting bovine immunoglobulin G is completed in 19 min with a limit of detection of 3.8 nM. The ultrafast analysis time and superior sensitivity demonstrated by this microfluidic HEI is promising for being used to develop the next-generation immunosensors.

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Acknowledgments

This work was supported by the National Science Foundation (NSF-OISE-0530203) and the NSF EPSCoR Fellowship (to P. Li). We gratefully thank Prof. Stephanus Büttgenbach at the Institute for Microtechnology of Technical University of Braunschweig for helpful research collaborations, Paul W. Johnson at the RI Genomics and Sequencing Center for his excellent technical assistance with the fluorescence imaging, and Kurt Broderick at the Microsystems Technology Laboratories of Massachusetts Institute of Technology for his valuable technical assistance with the microfluidic chip fabrication.

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Authors and Affiliations

  1. Department of Mechanical Engineering and Applied Mechanics, University of Rhode Island, Kingston, RI, 02881, USA

    Peng Li, Constantine Anagnostopoulos & Mohammad Faghri

  2. Food Science Department, Faculty of Agriculture, Ain Shams University, Cairo, 11241, Egypt

    Assem Abolmaaty

  3. Department of Biological Sciences, University of Rhode Island, Kingston, RI, 02881, USA

    Christine D’Amore

  4. Institute for Microtechnology, Technical University of Braunschweig, 38124, Braunschweig, Germany

    Stefanie Demming

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  1. Peng Li
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  2. Assem Abolmaaty
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  3. Christine D’Amore
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  4. Stefanie Demming
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  5. Constantine Anagnostopoulos
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  6. Mohammad Faghri
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Correspondence to Mohammad Faghri.

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Li, P., Abolmaaty, A., D’Amore, C. et al. Development of an ultrafast quantitative heterogeneous immunoassay on pre-functionalized poly(dimethylsiloxane) microfluidic chips for the next-generation immunosensors. Microfluid Nanofluid 7, 593 (2009). https://doi.org/10.1007/s10404-009-0422-9

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  • DOI: https://doi.org/10.1007/s10404-009-0422-9

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