Lanthanide-controlled protein switches: development and in vitro and in vivo applications.

Lanthanides, which are part of the rare earth elements group have numerous applications in electronics, medicine and energy storage.  However, our ability to extract them is not meeting the rapidly increasing demand. The discovery of the bacterial periplasmic lanthanide-binding protein lanmodulin spurred significant interest in developing biotechnological routes for lanthanide detection and extraction.

pH-Driven Enzymatic Breakdown and Release of Catalase from Alginate Hydrogel.

Stimuli-induced release resulting in biochemical transformations has received a lot of attention due to its application in controlled drug release. In this work, catalase (EC 1.11.

Enzymatically Triggered Drug Release from Microgels Controlled by Glucose Concentration.

This study aims to design microgels for controlled drug release via enzymatically generated pH changes in the presence of glucose. Modern medicine is focused on developing smart delivery systems with controlled release capabilities. In response to this demand, we present the synthesis, characterization, and enzymatically triggered drug release behavior of microgels based on poly(acrylic acid) modified with glucose oxidase (GOx) (p(AA-BIS)-GOx).

Chemical modification of α-chymotrypsin enabling its release from alginate hydrogel by electrochemically generated local pH change.

This study investigates the controlled release of α-chymotrypsin from an alginate hydrogel matrix. When protein molecules entrapped in the hydrogel matrix have a size smaller than the hydrogel pores, their hold/release from the polymer matrix are controlled by the electrostatic interaction between the guest molecules and host polymer. α-Chymotrypsin, as a model protein, was chemically modified with negatively charged species to change its pI and to convert its attractive interaction with a negatively charged alginate hydrogel matrix to a repulsion interaction allowing its release by pH-triggered signal.

Time-Separated Pulse Release-Activation of an Enzyme from Alginate-Polyethylenimine Hydrogels Using Electrochemically Generated Local pH Changes.

β-Glucosidase (EC 3.2.1.

Electrochemical and Biocatalytic Signal-Controlled Payload Release from a Metal-Organic Framework.

A metal-organic framework (MOF), ZIF-8, which is stable at neutral and slightly basic pH values in aqueous solutions and destabilized/dissolved under acidic conditions, is loaded with a pH-insensitive fluorescent dye, rhodamine-B isothiocyanate, as a model payload species. Then, the MOF species are immobilized at an electrode surface. The local (interfacial) pH value is rapidly decreased by means of an electrochemically stimulated ascorbate oxidation at +0.

Biosensors: Electrochemical Devices-General Concepts and Performance.

This review provides a general overview of different biosensors, mostly concentrating on electrochemical analytical devices, while briefly explaining general approaches to various kinds of biosensors, their construction and performance. A discussion on how all required components of biosensors are brought together to perform analytical work is offered. Different signal-transducing mechanisms are discussed, particularly addressing the immobilization of biomolecular components in the vicinity of a transducer interface and their functional integration with electronic devices.

Fluorometric biosensing of α-amylase using an artificial allosteric biosensor immobilized on nanostructured interface.

Protein biosensors hold a promise to transform the way we collect physiological data by enabling quantification of biomarkers outside of specialized laboratory environment. However, achieving high specificity and sensitivity in homogeneous assay format remains challenging. Here we report construction of fluorescent biosensor arrays based on artificial allosteric α-amylase-activated PQQ-dependent glucose dehydrogenase (Amy-GDH).

Stimulation-Inhibition of Protein Release from Alginate Hydrogels Using Electrochemically Generated Local pH Changes.

The electrochemically controlled release of proteins was studied in a Ca-cross-linked alginate hydrogel deposited on an electrode surface. The electrochemical oxidation of ascorbate or reduction of O was achieved upon applying electrical potentials +0.6 or -0.

Multifunctional Hybrid Nanocomposite Hydrogel Releasing Different Biomolecular Species Triggered with Different Biochemical Signals Processed by Orthogonal Biocatalytic Reactions.

A micro/nanoshaped system composed of alginate microspheres (microgels) decorated with silica oxide nanoparticles functionalized with nitroavidin was used for on-demand biomolecule release stimulated by different input signals. Enzymes preloaded in the microgels processed the applied signals producing either basic pH locally near the microspheres or generating HO inside the hydrogel, or both simultaneously. The pH increase resulted in cleavage of the affinity bonds between nitroavidin and biotin, then releasing the latter.

Biosensor Based on Peroxidase-Mimetic Nanozyme and Lactate Oxidase for Accurate L-Lactate Analysis in Beverages.

Precision analysis of the key biological metabolites such as L-lactate has great practical importance for many technological processes in food technology, including beverage production. Here we describe a new, highly selective, and sensitive biosensor for accurate L-lactate assay based on a combination of peroxidase-mimetic nanozymes with microbial lactate oxidase (LOx) immobilized onto the surface of a graphite-rod electrode (GE). The peroxidase-like nanozymes were synthesized using the debris of carbon microfibers (CFs) functionalized with hemin (H) and modified with gold nanoparticles (AuNPs) or platinum microparticles (PtMPs).

UV-Cured Green Polymers for Biosensorics: Correlation of Operational Parameters of Highly Sensitive Biosensors with Nano-Volumes and Adsorption Properties.

The investigated polymeric matrixes consisted of epoxidized linseed oil (ELO), acrylated epoxidized soybean oil (AESO), trimethylolpropane triglycidyl ether (RD1), vanillin dimethacrylate (VDM), triarylsulfonium hexafluorophosphate salts (PI), and 2,2-dimethoxy-2-phenylacetophenone (DMPA). Linseed oil-based (ELO/PI, ELO/10RD1/PI) and soybean oil-based (AESO/VDM, AESO/VDM/DMPA) polymers were obtained by cationic and radical photopolymerization reactions, respectively. In order to improve the cross-linking density of the resulting polymers, 10 mol.

Fluorescent sensor based on pyrroloquinoline quinone (PQQ)-glucose dehydrogenase for glucose detection with smartphone-adapted signal analysis.

A new nano-structured platform for fluorescent analysis using PQQ-dependent glucose dehydrogenase (PQQ-GDH) was developed, particularly using a smartphone for transduction and quantification of optical signals. The PQQ-GDH enzyme was immobilized on SiO nanoparticles deposited on glass microfiber filter paper, providing a high load of the biocatalytic enzyme. The platform was tested and optimized for glucose determination using a wild type of the PQQ-GDH enzyme.

Multiple pH waves generated electrochemically and propagated from an electrode surface.

Local pH changes were produced upon electrochemical reactions. Cyclic application of reductive and oxidative potentials resulted in the formation of pH waves in the form of distinct solution layers. Multiple layers with basic and acidic pH values were visualized with a fluorescence confocal microscope following fluorescence of pH-dependent dyes.

Electrochemically stimulated protein release from pH-switchable electrode-immobilized nitroavidin-biotin and avidin-iminobiotin systems.

Bovine serum albumin (BSA), used as a model protein, was immobilized on a buckypaper electrode by formation of covalent bonds with avidin/iminobiotin or nitroavidin/biotin complexes. pH-sensitive affinity interactions between avidin and iminobiotin or between nitroavidin and biotin allowed splitting of the affinity bonds upon pH variation, thus resulting in BSA release. Local (interfacial) pH was changed electrochemically.

Improvement of laccase biosensor characteristics using sulfur-doped TiO nanoparticles.

The search for new nanoscale materials with predictable properties to target the timely and fast detection of toxic components in wastewater is one of the most promising directions of modern biosensorics. We have shown that TiO nanoparticles modified with sulfur significantly improve the main operational parameters of laccase-based electrodes when compared with controls. The nanoparticle samples were labeled as TiOS(0.

A Universal Multichannel Platform for Assembling Enzyme-Based Boolean Logic Gates.

Concatenated enzyme-based Boolean logic gates activated with 5 chemical input signals were analyzed with a smartphone photo camera. Simultaneous detection of 32 input combinations was conveniently performed using enzyme-modified fiberglass sensing spots generating fluorescence with different intensities for the 0 and 1 binary outputs. The developed technology offers an easy readout method for multi-channel logic systems.

Nanostructured Interface Loaded with Chimeric Enzymes for Fluorimetric Quantification of Cyclosporine A and FK506.

Advances in protein engineering resulted in increased efforts to create protein biosensors that can replace instrumentation-heavy analytical and diagnostic methods. Sensitivity, amenability to multiplexing, and manufacturability remain to be among the key issues preventing broad utilization of protein biosensors. Here, we attempt to address these by constructing arrays utilizing protein biosensors based on the artificial allosteric variant of PQQ-glucose dehydrogenase (GDH).

Electrochemically switchable and tunable luciferase bioluminescence.

The biocatalytic activity of electrode-immobilized luciferase followed by bioluminescence emitted from the electrode surface was reversibly tuned and switched by applying electrochemical signals. When a reductive potential (-0.9 V vs.

Electrochemically produced local pH changes stimulating (bio)molecule release from pH-switchable electrode-immobilized avidin-biotin systems.

Immobilized avidin-biotin complexes were used to release biotinylated (bio)molecules upon producing local pH changes near an electrode surface by electrochemical reactions. The nitro-avidin complex with biotin was dissociated by increasing local pH with electrochemical O reduction. The avidin complex with iminobiotin was split by decreasing local pH with electrochemical oxidation of ascorbate.

A universal nanostructured bioanalytical platform for NAD-dependent enzymes based on the fluorescent output reading with a smartphone.

A nanostructured surface composed of SiO nanoparticles (200 nm) deposited on fiberglass was used as a platform for immobilization of NAD-dependent enzymes (alcohol and glucose dehydrogenases). The surface functionalized with the enzymes was further loaded with NAD(P)cofactor and 1-methoxy-5-methylphenazinium methyl sulfate (MPMS) redox species. The designed biocatalytic system was used for fluorescent analysis of substrates (ethanol and glucose used as example analyte substances).

A magneto-controlled biocatalytic cascade with a fluorescent output.

A biocatalytic cascade based on concerted operation of pyruvate kinase and luciferase with a bioluminescent output was switched reversibly between low and high activity by applying an external magnetic field at different positions or removing it. The enzymes participating in the reaction cascade were bound to magnetic nanoparticles to allow their translocation or aggregation/dispersion to be controlled by the magnetic field. The reaction intensity, measured as the bioluminescent output, was dependent on the effective distances between the enzymes transported on the magnetic nanoparticles controlled by the magnets.

"Smart" Delivery of Monoclonal Antibodies from a Magnetic Responsive Microgel Nanocomposite.

"Smart" drug-delivery systems have significant potential to increase therapeutic efficiency, avoid undesired immune responses, and minimize drug side effects. Herein, we report on an innovative strategy to control the drug release process using two magneto-activated materials operating in the system. One of them, a polyvinyl alcohol (PVA)-diboronate (DB)-interpenetrated (IPN) alginate (Alg) microgel nanocomposite (PVA-DB-IPN-Alg) loaded with magnetic nanoparticles (MNPs), is acting as a drug-delivery system.

Design of a methotrexate-controlled chemical dimerization system and its use in bio-electronic devices.

Natural evolution produced polypeptides that selectively recognize chemical entities and their polymers, ranging from ions to proteins and nucleic acids. Such selective interactions serve as entry points to biological signaling and metabolic pathways. The ability to engineer artificial versions of such entry points is a key goal of synthetic biology, bioengineering and bioelectronics.