Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Tim Wendlandt
,
Claudia Koch
,
Beate Britz
,
Anke Liedek
,
Nora Schmidt
,
Stefan Werner
ORCID logo ,
Yuri Gleba
,
Farnoosh Vahidpour
,
Melanie Welden
,
Arshak Poghossian
,
Michael J. Schöning
ORCID logo ,
Fabian J. Eber
,
Holger Jeske
,
Christina Wege
* ORCID logo
Version 1 : Received: 4 August 2023 / Approved: 7 August 2023 / Online: 7 August 2023 (12:23:31 CEST)

A peer-reviewed article of this Preprint also exists.

Wendlandt, T.; Koch, C.; Britz, B.; Liedek, A.; Schmidt, N.; Werner, S.; Gleba, Y.; Vahidpour, F.; Welden, M.; Poghossian, A.; Schöning, M.J.; Eber, F.J.; Jeske, H.; Wege, C. Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System. Viruses 2023, 15, 1951. Wendlandt, T.; Koch, C.; Britz, B.; Liedek, A.; Schmidt, N.; Werner, S.; Gleba, Y.; Vahidpour, F.; Welden, M.; Poghossian, A.; Schöning, M.J.; Eber, F.J.; Jeske, H.; Wege, C. Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System. Viruses 2023, 15, 1951.

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 wildtype 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 by a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as target, yielding synthetic multienzyme-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 achieved also on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable, TVCVPA-assisted electrochemical biosensing through modularly IgG-assembled sensor enzymes.

Keywords

turnip vein clearing virus (TVCV); tobacco mosaic virus (TMV); tobamovirus; enzyme nanocarrier; protein A; antibody; enzyme cascade; glucose oxidase (GOx); horseradish peroxidase (HRP); biosensor

Subject

Biology and Life Sciences, Virology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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