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Composable Microfluidic Plates (cPlate): A Simple and Scalable Fluid Manipulation System for Multiplexed Enzyme-Linked Immunosorbent Assay (ELISA)

  • Ziyi He
    Ziyi He
    C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, United States
    More by Ziyi He
  • Justin Huffman
    Justin Huffman
    C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, United States
  • Kathrine Curtin
    Kathrine Curtin
    Department of Mechanical and Aerospace Engineering, West Virginia University, Morgantown, West Virginia 26506, United States
  • Krista L. Garner
    Krista L. Garner
    Department of Physiology and Pharmacology, West Virginia University, Morgantown, West Virginia 26506, United States
    Center for Inhalation Toxicology, West Virginia University, Morgantown, West Virginia 26506, United States
  • Elizabeth C. Bowdridge
    Elizabeth C. Bowdridge
    Department of Physiology and Pharmacology, West Virginia University, Morgantown, West Virginia 26506, United States
    Center for Inhalation Toxicology, West Virginia University, Morgantown, West Virginia 26506, United States
  • Xiaojun Li
    Xiaojun Li
    C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, United States
    More by Xiaojun Li
  • Timothy R. Nurkiewicz
    Timothy R. Nurkiewicz
    Department of Physiology and Pharmacology, West Virginia University, Morgantown, West Virginia 26506, United States
    Center for Inhalation Toxicology, West Virginia University, Morgantown, West Virginia 26506, United States
  • , and 
  • Peng Li*
    Peng Li
    C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, United States
    *Email: [email protected]
    More by Peng Li
Cite this: Anal. Chem. 2021, 93, 3, 1489–1497
Publication Date (Web):December 16, 2020
https://doi.org/10.1021/acs.analchem.0c03651
Copyright © 2020 American Chemical Society

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    Abstract

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    Enzyme-linked immunosorbent assay (ELISA) is the gold standard method for protein biomarkers. However, scaling up ELISA for multiplexed biomarker analysis is not a trivial task due to the lengthy procedures for fluid manipulation and high reagent/sample consumption. Herein, we present a highly scalable multiplexed ELISA that achieves a similar level of performance to commercial single-target ELISA kits as well as shorter assay time, less consumption, and simpler procedures. This ELISA is enabled by a novel microscale fluid manipulation method, composable microfluidic plates (cPlate), which are comprised of miniaturized 96-well plates and their corresponding channel plates. By assembling and disassembling the plates, all of the fluid manipulations for 96 independent ELISA reactions can be achieved simultaneously without any external fluid manipulation equipment. Simultaneous quantification of four protein biomarkers in serum samples is demonstrated with the cPlate system, achieving high sensitivity and specificity (∼ pg/mL), short assay time (∼1 h), low consumption (∼5 μL/well), high scalability, and ease of use. This platform is further applied to probe the levels of three protein biomarkers related to vascular dysfunction under pulmonary nanoparticle exposure in rat’s plasma. Because of the low cost, portability, and instrument-free nature of the cPlate system, it will have great potential for multiplexed point-of-care testing in resource-limited regions.

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.0c03651.

    • Experimental details, cPlate device design, loading pattern, performance of uncoated/patterned well plates, volume consistency after washing, response curves for plasma proteins, correlations between cPlate and 96-well ELISA, and comparison between conventional ELISA and cPlate ELISA (PDF)

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