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

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TitleBiofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-tag.
Publication TypeJournal Article
Year of Publication2009
AuthorsWu, H-C, Shi, X-W, Tsao, C-Y, Lewandowski, AT, Fernandes, R, Hung, C-W, DeShong, P, Kobatake, E, Valdes, JJ, Payne, GF, Bentley, WE
JournalBiotechnol Bioeng
Volume103
Issue2
Pagination231-40
Date Published2009 Jun 1
ISSN1097-0290
KeywordsAmino Acid Motifs, Antibodies, Antigens, Binding Sites, Biotechnology, Chitosan, Hydrogen-Ion Concentration, Macromolecular Substances, Protein Binding, Recombinant Fusion Proteins
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.

DOI10.1002/bit.22238
Alternate JournalBiotechnol. Bioeng.
PubMed ID19224560