TY  - JOUR
A1  - Wendlandt, Tim
A1  - Koch, Claudia
A1  - Britz, Beate
A1  - Liedek, Anke
A1  - Schmidt, Nora
A1  - Werner, Stefan
A1  - Gleba, Yuri
A1  - Vahidpour, Farnoosh
A1  - Welden, Melanie
A1  - Poghossian, Arshak
A1  - Schöning, Michael Josef
T1  - Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System
JF  - Viruses
N2  - Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.
KW  - biosensor
KW  - horseradish peroxidase (HRP)
KW  - glucose oxidase (GOx)
KW  - enzyme cascade
KW  - turnip vein clearing virus (TVCV)
KW  - tobacco mosaic virus (TMV)
Y1  - 2023
U6  - http://dx.doi.org/doi.org/10.3390/v15091951
SN  - 1999-4915
N1  - This article belongs to the Special Issue "Tobamoviruses 2023"
VL  - 9
IS  - 15
PB  - MDPI
CY  - Basel
ER  -