Common Causes of Glucose Oxidase Instability in <i>In Vivo</i> Biosensing: A Brief Review

Harris, James M.; Reyes, Catherine D.; López, Gabriel P.

doi:10.1177/193229681300700428

Cited by 92 publications

(63 citation statements)

References 78 publications

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“…It is possible that the presence of the Aga2 protein at the N-terminus of GOx affects its enzymatic activity. Alternatively, the higher surface density of GOx in case of pCTCON may result in lower observed activity due to local depletion of oxygen or inactivation of GOx due to local accumulation of hydrogen peroxide, as observed in the case of sensors employing immobilized GOx 34–36 . However, further investigation into the apparent decrease in enzymatic activity of surface displayed GOx when the pCTCON vector is used is outside the scope of this study.…”

Section: Resultsmentioning

confidence: 99%

Inefficient Ribosomal Skipping Enables Simultaneous Secretion and Display of Proteins in Saccharomyces cerevisiae

et al. 2017

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The need for recombinant expression of soluble protein slows down validation of engineered proteins isolated from combinatorial libraries, and limits the number of protein variants evaluated. To overcome this bottleneck, we describe a system based on inefficient ribosomal skipping, for simultaneous cell surface display and soluble secretion of proteins in Saccharomyces cerevisiae. Ribosomal skipping mediated by “self-cleaving” 2A peptides produces two proteins from a single open reading frame. Incorporation of the F2A peptide sequence – with ~ 50% efficiency of ribosomal skipping – between the protein of interest and the yeast cell wall protein Aga2 results in simultaneous expression of both solubly secreted protein, and the protein-Aga2 fusion that is tethered to the yeast cell surface. We show that binding proteins derived from the Sso7d scaffold and the homodimeric enzyme glucose oxidase (GOx) can be simultaneously secreted solubly, and expressed as yeast cell surface fusions, using the F2A-based system. Further, a combinatorial library of Sso7d mutants can be screened to isolate binders with higher affinity for a model target (lysozyme), and the pool of higher affinity binders can be characterized in soluble form. Significantly, we show that both N- and C-terminal fusions to Aga2 can be simultaneously secreted solubly and displayed on the cell surface; this is particularly advantageous because protein functionality can be affected by the specific position of Aga2 in the protein fusion. We expect that the F2A-based yeast surface display and secretion system will be a useful tool for protein engineering and enable efficient characterization of individual clones isolated from combinatorial libraries.

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Section: Resultsmentioning

confidence: 99%

Inefficient Ribosomal Skipping Enables Simultaneous Secretion and Display of Proteins in Saccharomyces cerevisiae

et al. 2017

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“…The variation of these CV curves during continuous cycling indicated the CoHCF!CoO x transformation process involved different and simultaneous electrochemical process on NPG surface. The initial process involved the transformation of CoHCF to more stable Co(OH) 2 Fig. 1C.…”

Section: Preparation and Electrochemical Characterization Of Coo X /Nmentioning

confidence: 99%

“…One is based on enzymes that can specifically catalyze glucose (enzymatic glucose sensors) and the other is based on the direct electrooxidization of glucose on electrode materials (nonenzymatic glucose sensors). Enzymatic glucose sensors are easily affected by environmental changes due to the inherent drawbacks of enzymes and the immobilization procedures are usually time-consuming [1,2]. Therefore, much effort has been devoted to the development of enzyme-free glucose sensors with excellent analytical performance.…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Fabrication of Cobalt Oxides/Nanoporous Gold Composite Electrode and its Nonenzymatic Glucose Sensing Performance

Zhou

Tang²,

Xia

et al. 2016

Electroanalysis

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A nonenzymatic glucose sensor was successfully established by electrochemically decorating cobalt oxides (CoOx) on a nanoporous gold electrode (NPG) using cobalt hexacyanoferrate (CoHCF) as a precursor. It exhibited high sensitivity and long‐term stability as well as satisfactory quantification of glucose concentration in human serum samples. The morphology and surface analysis of the resulting CoOx/NPG were carefully characterized. Two detection methods, cyclic voltammetry and amperometry, were employed to evaluate the performance of CoOx/NPG towards glucose sensing in alkaline solution. Using cyclic voltammetry, at −0.5 V, the glucose partial oxidation peak current is linear to the glucose concentration up to 14 mM with a sensitivity of 283.7 µA mM−1 cm−2. A linear amperometric response at 0.55 V was obtained in the glucose concentration range from 2 µM to 2 mM with a sensitivity of 2025 µA mM−1 cm−2 and a response time <3 s.

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“…Unfortunately, many factors may degrade the glucose oxidase structure following by a decrease of sensor activity. Hydrogen peroxide, unstable temperature, blood low molecular weight materials, and interstitial fluids are the major reasons for biosensor instability (Harris, Reyes, and Lopez 2013). Hydrogen peroxide produced during reaction catalyzed by glucose oxidase is the most frequent factor of glucose oxidase degradation (Ferri, Kojima, and Sode 2011).…”

Section: Biorecognition Elements Of Glucose Biosensorsmentioning

confidence: 99%

Biosensors for Blood Glucose and Diabetes Diagnosis: Evolution, Construction, and Current Status

Martinková¹,

Pohanka²

2015

Analytical Letters

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Diabetes mellitus is a worldwide health problem and complications associated with the disease are a significant cause of death in the world. Monitoring the blood glucose level is the first step during diagnosis of diabetes mellitus; hence rapid and accurate method of diagnosis is necessary for prevention of lethal complications. Current research is directed towards miniaturization of analytical equipment and also towards a decrease in consumption of biological materials and chemical substances. As a result, biosensors are available for sensitive, accurate, and low cost measurements. The first glucose enzymatic biosensor was proposed by Clark and Lyons (1962).Many different applications and innovations of biosensor have been performed since this work. Biosensors consist of a biorecognition element and physicochemical transducer providing aDownloaded by [New York University] at 01:28 22 June 20152 measurable signal. In case of glucose biosensors, enzyme glucose oxidase is used as a biorecognition element (or biotransducer in some sources) converting glucose to gluconic acid.An electrochemical device was the most common physicochemical transducer for many years, but the optical transducers are becoming more common now. In spite of the advantages of enzymatic glucose biosensors, many disadvantages also were encountered. Consequently, some researchers developed non-enzymatic glucose biosensors. This review covers evolution of glucose biosensors, construction of traditional and marketed types of biosensors, and promising applications.

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Common Causes of Glucose Oxidase Instability in In Vivo Biosensing: A Brief Review

Cited by 92 publications

References 78 publications

Inefficient Ribosomal Skipping Enables Simultaneous Secretion and Display of Proteins in Saccharomyces cerevisiae

Inefficient Ribosomal Skipping Enables Simultaneous Secretion and Display of Proteins in Saccharomyces cerevisiae

Electrochemical Fabrication of Cobalt Oxides/Nanoporous Gold Composite Electrode and its Nonenzymatic Glucose Sensing Performance

Biosensors for Blood Glucose and Diabetes Diagnosis: Evolution, Construction, and Current Status

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