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

Biotechnol Bioeng. 2009 Jun 1;103(2):231-40. doi: 10.1002/bit.22238.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Motifs
  • Antibodies / genetics*
  • Antibodies / isolation & purification
  • Antibodies / metabolism*
  • Antigens / genetics*
  • Antigens / isolation & purification
  • Antigens / metabolism*
  • Binding Sites
  • Biotechnology / methods
  • Chitosan / metabolism*
  • Hydrogen-Ion Concentration
  • Macromolecular Substances*
  • Protein Binding
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism

Substances

  • Antibodies
  • Antigens
  • Macromolecular Substances
  • Recombinant Fusion Proteins
  • Chitosan