Hsuan-Chen Wu and Xiao-Wen Shi contributed equally to this work.
Biofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-tag†
Article first published online: 30 DEC 2008
Copyright © 2008 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 103, Issue 2, pages 231–240, 1 June 2009
How to Cite
Wu, H.-C., Shi, X.-W., Tsao, C.-Y., Lewandowski, A. T., Fernandes, R., Hung, C.-W., DeShong, P., Kobatake, E., Valdes, J. J., Payne, G. F. and Bentley, W. E. (2009), Biofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-tag. Biotechnol. Bioeng., 103: 231–240. doi: 10.1002/bit.22238
- Issue published online: 21 APR 2009
- Article first published online: 30 DEC 2008
- Accepted manuscript online: 30 DEC 2008 12:00AM EST
- Manuscript Accepted: 15 DEC 2008
- Manuscript Revised: 4 DEC 2008
- Manuscript Received: 25 AUG 2008
- US Army DTRA
- R.W. Deutsch Foundation
- National Science Foundation. Grant Numbers: CBET-0650650, EFRI-0735987
- protein G;
- tyrosine tag
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.