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Keywords
- Label-free detection (3)
- capacitive field-effect sensor (3)
- field-effect sensor (3)
- tobacco mosaic virus (TMV) (3)
- Capacitive field-effect sensor (2)
- Field-effect sensor (2)
- LAPS (2)
- gold nanoparticles (2)
- (Bio)degradation (1)
- CNOT (1)
- Capacitive field-effect (1)
- Capacitive model (1)
- C–V method (1)
- DNA biosensor (1)
- DNA hybridization (1)
- Electrolyte–insulator–semiconductor (1)
- Enzyme coverage (1)
- Enzyme logic gate (1)
- Field effect (1)
- Field-effect biosensor (1)
- Gold nanoparticles (1)
- Impedance spectroscopy (1)
- Layer-by-layer adsorption (1)
- Multianalyte detection (1)
- Multicell (1)
- Multiplexing (1)
- Penicillin (1)
- Plant virus (1)
- Poly(allylamine hydrochloride) (1)
- Poly(d,l-lacticacid) (1)
- Real-time monitoring (1)
- TMV adsorption (1)
- Ta₂O₅ gate (1)
- Tobacco mosaic virus (TMV) (1)
- XOR (1)
- Zeta potential (1)
- aminooctanethiol (1)
- atomic layer deposition (1)
- barium strontium titanate (1)
- bi-enzyme biosensor (1)
- biosensor (1)
- capacitive EIS sensor (1)
- capacitive field-effect sensors (1)
- capacitive model (1)
- contactless conductivity sensor (1)
- control gate (1)
- detection of charged macromolecules (1)
- electrolyte-insulator-semiconductor capacitors (1)
- enzymatic (bio)degradation (1)
- enzyme cascade (1)
- enzyme-logic gate (1)
- equivalent circuit (1)
- glucose oxidase (GOx) (1)
- high-k material (1)
- horseradish peroxidase (HRP) (1)
- hydrogen peroxide (1)
- impedance spectroscopy (1)
- in-situ monitoring (1)
- lable-free detection (1)
- multi-functional material (1)
- multianalyte detection (1)
- nanoparticle coverage (1)
- on-chip integrated addressable EISCAP sensors (1)
- pH sensors (1)
- penicillinase (1)
- plant virus detection (1)
- poly(d, l-lactic acid) (1)
- polystyrene sulfonate (1)
- turnip vein clearing virus (TVCV) (1)
- ultrathin gate insulators (1)
- urease (1)
Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, tobacco mosaic virus (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO₂-Ta₂O₅ layer structure for the sequential detection of penicillin and urea. The TMV-assisted bi-enzyme EISCAP biosensor exhibited a high urea and penicillin sensitivity of 54 and 85 mV/dec, respectively, in the concentration range of 0.1–3 mM. For comparison, the characteristics of single-enzyme EISCAP biosensors modified with TMV particles immobilized with either penicillinase or urease were also investigated. The surface morphology of the TMV-modified Ta₂O₅-gate was analyzed by scanning electron microscopy. Additionally, the bi-enzyme EISCAP was applied to mimic an XOR (Exclusive OR) enzyme logic gate.
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.
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.
Electrolyte-insulator-semiconductor capacitors (EISCAP) belong to field-effect sensors having an attractive transducer architecture for constructing various biochemical sensors. In this study, a capacitive model of enzyme-modified EISCAPs has been developed and the impact of the surface coverage of immobilized enzymes on its capacitance-voltage and constant-capacitance characteristics was studied theoretically and experimentally. The used multicell arrangement enables a multiplexed electrochemical characterization of up to sixteen EISCAPs. Different enzyme coverages have been achieved by means of parallel electrical connection of bare and enzyme-covered single EISCAPs in diverse combinations. As predicted by the model, with increasing the enzyme coverage, both the shift of capacitance-voltage curves and the amplitude of the constant-capacitance signal increase, resulting in an enhancement of analyte sensitivity of the EISCAP biosensor. In addition, the capability of the multicell arrangement with multi-enzyme covered EISCAPs for sequentially detecting multianalytes (penicillin and urea) utilizing the enzymes penicillinase and urease has been experimentally demonstrated and discussed.