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Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System

  • 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.

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Author:Tim Wendlandt, Claudia Koch, Beate Britz, Anke Liedek, Nora Schmidt, Stefan Werner, Yuri Gleba, Farnoosh Vahidpour, Melanie Welden, Arshak Poghossian, Michael Josef SchöningORCiD
Parent Title (English):Viruses
Place of publication:Basel
Document Type:Article
Year of Completion:2023
Date of first Publication:2023/09/19
Date of the Publication (Server):2023/09/21
Tag:biosensor; enzyme cascade; glucose oxidase (GOx); horseradish peroxidase (HRP); tobacco mosaic virus (TMV); turnip vein clearing virus (TVCV)
First Page:Artikel 1951
This article belongs to the Special Issue "Tobamoviruses 2023"
Institutes:FH Aachen / INB - Institut für Nano- und Biotechnologien
collections:Verlag / MDPI
Licence (German):License LogoCreative Commons - Namensnennung