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http://dx.doi.org/10.18419/opus-13895
Autor(en): | Wendlandt, Tim Koch, Claudia Britz, Beate Liedek, Anke Schmidt, Nora Werner, Stefan Gleba, Yuri Vahidpour, Farnoosh Welden, Melanie Poghossian, Arshak Schöning, Michael J. Eber, Fabian J. Jeske, Holger Wege, Christina |
Titel: | Facile purification and use of tobamoviral nanocarriers for antibody-mediated display of a two-enzyme system |
Erscheinungsdatum: | 2023 |
Dokumentart: | Zeitschriftenartikel |
Seiten: | 32 |
Erschienen in: | Viruses 15 (2023), No.1951 |
URI: | http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-ds-139147 http://elib.uni-stuttgart.de/handle/11682/13914 http://dx.doi.org/10.18419/opus-13895 |
ISSN: | 1999-4915 |
Zusammenfassung: | 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. |
Enthalten in den Sammlungen: | 04 Fakultät Energie-, Verfahrens- und Biotechnik |
Dateien zu dieser Ressource:
Datei | Beschreibung | Größe | Format | |
---|---|---|---|---|
viruses-15-01951.pdf | Artikel | 4,36 MB | Adobe PDF | Öffnen/Anzeigen |
viruses-15-01951-s001.zip | Supplement | 513,09 kB | Unknown | Öffnen/Anzeigen |
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