title: Biofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-tag.
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.
authors.affiliation: Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742, USA.
authors.forename: Hsuan-Chen
authors.initials: HC
authors.lastname: Wu
authors.forename: Xiao-Wen
authors.initials: XW
authors.lastname: Shi
authors.forename: Chen-Yu
authors.initials: CY
authors.lastname: Tsao
authors.forename: Angela T
authors.initials: AT
authors.lastname: Lewandowski
authors.forename: Rohan
authors.initials: R
authors.lastname: Fernandes
authors.forename: Chi-Wei
authors.initials: CW
authors.lastname: Hung
authors.forename: Philip
authors.initials: P
authors.lastname: DeShong
authors.forename: Eiry
authors.initials: E
authors.lastname: Kobatake
authors.forename: James J
authors.initials: JJ
authors.lastname: Valdes
authors.forename: Gregory F
authors.initials: GF
authors.lastname: Payne
authors.forename: William E
authors.initials: WE
authors.lastname: Bentley
mesh_terms: D020816:Amino Acid Motifs; D000906:Antibodies / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000941:Antigens / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D001709:Biotechnology / Q000379:methods; D048271:Chitosan / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000906:Antibodies; D000941:Antigens; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D048271:Chitosan