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Activity of free and immobilized glucose oxidase: An electrochemical study

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Abstract

The activity of free and immobilized glucose oxidase was determined using a sandwich type thin-layer electrochemical cell. The thin-layer cell consisted of a gold electrode deposited on a glass microscope slide, 165 μm thick Teflon TFE spacers, and a glass cover. Enzyme activity was determined by using cyclic voltammetry to measure the amount of hydrogen peroxide produced in the glucose oxidase catalyzed redox reaction between glucose and oxygen in the thin-layer cell. The specific activity of 13.4 nM glucose oxidase in 0.2 M aqueous sodium phosphate, pH 5.2 at room temperature, was calculated to be 4.34 U/mg GOx. Under the same experimental conditions, qualitative detection of the activity of glucose oxidase covalently immobolized to a thin radiofrequency plasma modified poly(etherurethaneurea) film was demonstrated.

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Abbreviations

CDAP:

1-cyano-4-dimethylaminopyridinium tetrafluoroborate

DD water:

distilled, deionized water

F :

Farraday's constant

GOx:

glucose oxidase

GOx-PPNVP/PEUU:

glucose oxidase immobilized onto poly(etherurethaneurea) modified with plasma polymerizedN-vinyl-2-pyrrolidone

M :

number of moles

PEUU:

poly(etherurethaneurea)

PPNVP:

plasma polymerizedN-vinyl-2-pyrrolidone

PPNVP/PEUU:

poly(etherurethaneurea) modified with plasma polymerizedN-vinyl-2-pyrrolidone

Q :

charge

R :

resistance

RHE:

reversible hydrogen electrode

RIA:

radioimmunoassay

SCE:

saturated calomel electrode

t :

sampling time

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Authors and Affiliations

  1. Department of Biomedical Engineering, Case Western Reserve University, 44106, Cleveland, OH

    Michael J. Danilich & Roger E. Marchant

  2. Department of Chemistry, Case Western Reserve University, 44106, Cleveland, OH

    Dominic Gervasio

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  1. Michael J. Danilich
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  2. Dominic Gervasio
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  3. Roger E. Marchant
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Danilich, M.J., Gervasio, D. & Marchant, R.E. Activity of free and immobilized glucose oxidase: An electrochemical study. Ann Biomed Eng 21, 655–668 (1993). https://doi.org/10.1007/BF02368645

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