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. ThisImmunosorbent 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.…
Document Type: | Article (reviewed) |
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Zitierlink: | https://opus.hs-offenburg.de/8078 | Bibliografische Angaben |
Title (English): | Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System |
Author: | Tim Wendlandt, Claudia Koch, Beate Britz, Anke Liedek, Nora Schmidt, Stefan Werner, Yuri Gleba, Farnoosh Vahidpour, Melanie Welden, Arshak Poghossian, Michael J. Schöning, Fabian EberStaff MemberGND, Holger Jeske, Christina Wege |
Year of Publication: | 2023 |
Place of publication: | Basel |
Publisher: | MDPI |
First Page: | 1 |
Last Page: | 32 |
Article Number: | 1951 |
Parent Title (English): | Viruses |
Volume: | 15 |
Issue: | 9 |
ISSN: | 1999-4915 |
DOI: | https://doi.org/10.3390/v15091951 |
URN: | https://urn:nbn:de:bsz:ofb1-opus4-80787 |
Language: | English | Inhaltliche Informationen |
Institutes: | Fakultät Maschinenbau und Verfahrenstechnik (M+V) |
Institutes: | Bibliografie |
Tag: | biosensor; enzyme cascade; tobacco mosaic virus | Formale Angaben |
Relevance: | Wiss. Zeitschriftenartikel reviewed: Listung in Master Journal List |
Open Access: | Open Access |
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Licence (German): | Creative Commons - CC BY - Namensnennung 4.0 International |