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Biofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-tag

Authors

  • Hsuan-Chen Wu,

    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
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  • Xiao-Wen Shi,

    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
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  • Chen-Yu Tsao,

    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
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  • Angela T. Lewandowski,

    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
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  • Rohan Fernandes,

    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
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  • Chi-Wei Hung,

    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
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  • Philip DeShong,

    1. Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland
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  • Eiry Kobatake,

    1. Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan
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  • James J. Valdes,

    1. US Army Edgewood Chemical Biological Center, AMSRD-ECB-AP-B/Michel E3330, Aberdeen Proving Ground, Maryland
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  • Gregory F. Payne,

    Corresponding author
    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
    • Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075.
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  • William E. Bentley

    Corresponding author
    1. Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742; telephone: 1-301-405-8389; fax: 1-301-314-9075
    2. Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953
    • Fischell Department of Bioengineering, University of Maryland, College Park, Maryland 20742; telephone: 1-301-405-4321; fax: 1-301-405-9953.
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  • Hsuan-Chen Wu and Xiao-Wen Shi contributed equally to this work.

Abstract

We report the assembly of seven different antibodies (and two antigens) into functional supramolecular structures that are specifically designed to facilitate integration into devices using entirely biologically based bottom-up fabrication. This is enabled by the creation of an engineered IgG-binding domain (HG3T) with an N-terminal hexahistidine tag that facilitates purification and a C-terminal enzyme-activatable pentatyrosine “pro-tag” that facilitates covalent coupling to the pH stimuli-responsive polysaccharide, chitosan. Because we confer pH-stimuli responsiveness to the IgG-binding domain, it can be electrodeposited or otherwise assembled into many configurations. Importantly, we demonstrate the loading of both HG3T and antibodies can be achieved in a linear fashion so that quantitative assessment of antibodies and antigens is feasible. Our demonstration formats include: conventional multiwell plates, micropatterned electrodes, and fiber networks. We believe biologically based fabrication (i.e., biofabrication) provides bottom-up hierarchical assembly of a variety of nanoscale components for applications that range from point-of-care diagnostics to smart fabrics. Biotechnol. Bioeng. 2009;103: 231–240. © 2008 Wiley Periodicals, Inc.

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