Comparison of different methods of glucose oxidase immobilization

Valentová, Olga; Marek, M.; Švec, František; Štamberg, J.; Vodrážka, Z.

doi:10.1002/bit.260230913

Cited by 44 publications

(10 citation statements)

References 16 publications

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“…bIgG has a highly asymmetric shape, packing on a smooth gold surface at a density of ≈1.28 fg/μm 2 [122], corresponding to 13,000 molecules on a saturated microsensor. Numerous factors affect the packing densities achieved by the components during sequential binding of different molecules to form layers on a microsensor: the biosensor surface texture (sputtered-gold is very rough, unlike the evaporated-and-annealed-gold film on a typical SPR chip, which may enhance binding) [105], the method used to attach the first layer of biomolecules to the gold surface (e.g., crosslinkers, buffers, passivators) [118–121,123], and the binding affinities of the components.…”

Section: Resultsmentioning

confidence: 99%

Label-Free Microcavity Biosensors: Steps towards Personalized Medicine

Amarie

Glazier

2012

Sensors

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Personalized medicine has the potential to improve our ability to maintain health and treat disease, while ameliorating continuously rising healthcare costs. Translation of basic research findings to clinical applications within regulatory compliance is required for personalized medicine to become the new foundation for practice of medicine. Deploying even a few of the thousands of potential diagnostic biomarkers identified each year as part of personalized treatment workflows requires clinically efficient biosensor technologies to monitor multiple biomarkers in patients in real time. This paper discusses a critical component of a regulatory system, a microcavity optical biosensor for label-free monitoring of biomolecular interactions at physiologically-relevant concentrations. While most current biosensor research focuses on improving sensitivity, this paper emphasizes other characteristics a biosensor technology requires to be practical in a clinical setting, presenting robust microcavity biosensors which are easy to manufacture and integrate with microfluidics into flexible and redesignable platforms making the microcavity biosensors deployable for continuous monitoring of biomarkers in body fluids in the clinic, in dense 2D random arrays for high-throughput applications like drug-library screening in interactomics, and of the secretory behavior of single cells in the laboratory.

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Section: Resultsmentioning

confidence: 99%

Label-Free Microcavity Biosensors: Steps towards Personalized Medicine

Amarie

Glazier

2012

Sensors

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“…11,12 The goal of our recent investigations is the development of new ultrathin and transparent solid phases with an incorporated oxidoreductase enzyme for fiberoptical biosensors. To this end, a novel film-forming NH 2 -cellulose derivative, 6-deoxy-6-(4-aminophenyl)-amino)cellulosetosylate (NH 2 -cellulose film), 13 and a novel NH 2 -functionalized coating on glass (NH 2 -glass) 14 were developed as transparent NH 2 -functional support matrixes.…”

Section: Introductionmentioning

confidence: 99%

Stabilization of Activity of Oxidoreductases by Their Immobilization onto Special Functionalized Glass and Novel Aminocellulose Film Using Different Coupling Reagents

et al. 2002

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Glucose oxidase (GOD), horseradish peroxidase (HRP), and lactate oxidase (LOD) were covalently immobilized on special NH 2 -functionalized glass and on a novel NH 2 -cellulose film via 13 different coupling reagents. The properties of these immobilized enzymes, such as activity, storage stability, and thermostability, are strongly dependent on the coupling reagent. For example, GOD immobilized by cyanuric chloride on the NH 2 -cellulose film loses approximately half of its immobilized activity after 30 days of storage at 4°C or after treatment at 65°C for 30 min. In contrast, GOD immobilized by L-ascorbic acid onto the same NH 2 -cellulose film retains 90% of its initial activity after 1 year of storage at 4°C and 92% after heat treatment at 65°C for 30 min. Unlike GOD, in the case of LOD only immobilization on special NH 2 -functionalized glass, e.g., via cyanuric chloride, led to a stabilization of the enzyme activity in comparison to the native enzyme. The operational stability of immobilized HRP was up to 40 times higher than that of the native enzyme if coupling to the new NH 2 -cellulose film led to an amide or sulfonamide bond. Regarding the kinetics of the immobilized enzymes, the coupling reagent plays a minor role for the enzyme substrate affinity, which is characterized by the apparent Michaelis constant (K M,app ). The NH 2 -functionalized support material as well as the immobilized density of the protein and/or immobilized activity has a strong influence on the K M,app value. In all cases, K M,app decreases with increasing immobilized enzyme protein density and particularly drastically for GOD.

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“…10 and comprises aminolysis of the epoxide ring using ammonia or a diamine, followed by activation with dialdehyde most often glutaraldehyde [69]. The aldehyde functionality is then used for the reaction with an enzyme, and similarly to the previous technique, the labile imine double bond must be hydrogenated.…”

Section: Aminolysis Followed By Dialdehyde Activationmentioning

confidence: 99%

Less common applications of monoliths: I. Microscale protein mapping with proteolytic enzymes immobilized on monolithic supports

Švec

2006

Electrophoresis

131

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This review summarizes the recent contributions to the rapidly growing area of immobilized enzymes employing both silica and synthetic polymer-based monoliths as supports. Focus is mainly on immobilized proteolytic enzyme reactors designed for studies in proteomics. Porous monoliths emerged first as a new class of stationary phases for HPLC in the early 1990s. Soon thereafter, they were also used as supports for immobilization of proteins and preparation of both stationary phases for bioaffinity chromatography and enzymatic reactors. Organic polymer-based monoliths are typically prepared using a simple molding process carried out within the confines of a "mold" such as chromatographic column or capillary. Polymerization of a mixture comprising monomers, initiator, and porogenic solvent affords macroporous materials. In contrast, silica-based monoliths are first formed as a rigid rod from tetraalkoxysilane in the presence of PEG and subsequently encased with a plastic tube. Both types of monolith feature large through-pores that enable a rapid flow-through. Since all the solutions must flow through the monolith, the convection considerably accelerates mass transfer within the monolith. As a result, reactors including enzyme immobilized on monolithic support exhibit much higher activity compared to the reactions in solution.

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Comparison of different methods of glucose oxidase immobilization

Cited by 44 publications

References 16 publications

Label-Free Microcavity Biosensors: Steps towards Personalized Medicine

Label-Free Microcavity Biosensors: Steps towards Personalized Medicine

Stabilization of Activity of Oxidoreductases by Their Immobilization onto Special Functionalized Glass and Novel Aminocellulose Film Using Different Coupling Reagents

Less common applications of monoliths: I. Microscale protein mapping with proteolytic enzymes immobilized on monolithic supports

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