|Title||Biofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-tag.|
|Publication Type||Journal Article|
|Year of Publication||2009|
|Authors||Wu, 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|
|Date Published||2009 Jun 1|
|Keywords||Amino Acid Motifs, Antibodies, Antigens, Binding Sites, Biotechnology, Chitosan, Hydrogen-Ion Concentration, Macromolecular Substances, Protein Binding, Recombinant Fusion Proteins|
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.
|Alternate Journal||Biotechnol. Bioeng.|