Please use this identifier to cite or link to this item: http://dx.doi.org/10.18419/opus-13895
Authors: 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
Title: Facile purification and use of tobamoviral nanocarriers for antibody-mediated display of a two-enzyme system
Issue Date: 2023
metadata.ubs.publikation.typ: Zeitschriftenartikel
metadata.ubs.publikation.seiten: 32
metadata.ubs.publikation.source: 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
Abstract: 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.
Appears in Collections:04 Fakultät Energie-, Verfahrens- und Biotechnik

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