TY - JOUR A1 - Wendlandt, Tim A1 - Koch, Claudia A1 - Britz, Beate A1 - Liedek, Anke A1 - Schmidt, Nora A1 - Werner, Stefan A1 - Gleba, Yuri A1 - Vahidpour, Farnoosh A1 - Welden, Melanie A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System JF - Viruses N2 - 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. KW - biosensor KW - horseradish peroxidase (HRP) KW - glucose oxidase (GOx) KW - enzyme cascade KW - turnip vein clearing virus (TVCV) KW - tobacco mosaic virus (TMV) Y1 - 2023 U6 - http://dx.doi.org/doi.org/10.3390/v15091951 SN - 1999-4915 N1 - This article belongs to the Special Issue "Tobamoviruses 2023" VL - 9 IS - 15 PB - MDPI CY - Basel ER - TY - JOUR A1 - Karschuck, Tobias A1 - Schmidt, Stefan A1 - Achtsnicht, Stefan A1 - Poghossian, Arshak A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Multiplexing system for automated characterization of a capacitive field-effect sensor array JF - Physica Status Solidi A N2 - In comparison to single-analyte devices, multiplexed systems for a multianalyte detection offer a reduced assay time and sample volume, low cost, and high throughput. Herein, a multiplexing platform for an automated quasi-simultaneous characterization of multiple (up to 16) capacitive field-effect sensors by the capacitive–voltage (C–V) and the constant-capacitance (ConCap) mode is presented. The sensors are mounted in a newly designed multicell arrangement with one common reference electrode and are electrically connected to the impedance analyzer via the base station. A Python script for the automated characterization of the sensors executes the user-defined measurement protocol. The developed multiplexing system is tested for pH measurements and the label-free detection of ligand-stabilized, charged gold nanoparticles. KW - Capacitive field-effect sensor KW - Gold nanoparticles KW - Label-free detection KW - Multicell KW - Multiplexing Y1 - 2023 U6 - http://dx.doi.org/10.1002/pssa.202300265 SN - 1862-6300 (Print) SN - 1862-6319 (Online) N1 - Corresponding author: Michael Josef Schöning VL - 220 IS - 22 PB - Wiley-VCH CY - Weinheim ER -