Biofabrication of antibodies and antigens via IgG-binding domain engineered with activatable pentatyrosine pro-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.
Wu
Hsuan-Chen
HC
Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, Maryland 20742, USA.
Shi
Xiao-Wen
XW
Tsao
Chen-Yu
CY
Lewandowski
Angela T
AT
Fernandes
Rohan
R
Hung
Chi-Wei
CW
DeShong
Philip
P
Kobatake
Eiry
E
Valdes
James J
JJ
Payne
Gregory F
GF
Bentley
William E
WE
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
D000906:Antibodies; D000941:Antigens; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D048271:Chitosan