Wendlandt, TimKoch, ClaudiaBritz, BeateLiedek, AnkeSchmidt, NoraWerner, StefanGleba, YuriVahidpour, FarnooshWelden, MelaniePoghossian, ArshakSchöning, Michael J.Eber, Fabian J.Jeske, HolgerWege, Christina2024-01-292024-01-2920231999-49151879498251http://nbn-resolving.de/urn:nbn:de:bsz:93-opus-ds-139147http://elib.uni-stuttgart.de/handle/11682/13914http://dx.doi.org/10.18419/opus-13895Immunosorbent 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.eninfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/4.0/570610article2023-10-09