Colloidal gold as a biocompatible immobilization matrix suitable for the fabrication of enzyme electrodes by electrodeposition

Glucose oxidase, horseradish peroxidase, xanthine oxidase, and carbonic anhydrase have been adsorbed to colloidal gold sols with good retention of enzymatic activity. Adsorption of xanthine oxidase on colloidal gold did not result in a change in enzymatic activity as determined by active site titration with the stoichiometric inhibitor pterin aldehyde and by measurement of the apparent Michaelis constant (K′M). Gold sols with adsorbed glucose oxidase, horseradish peroxidase, and xanthine oxidase have also been electrodeposited onto conducting matrices (platinum gauze and/or glassy carbon) to make enzyme electrodes. These electrodes retained enzymatic activity and, more importantly, gave an electrochemical response to the enzyme substrate in the presence of an appropriate electron transfer mediator. Our results demonstrate the utility of colloidal gold as a biocompatible enzyme imobilization matrix suitable for the fabrication of enzyme electrodes. © 1992 John Wiley & Sons, Inc.

[1]  P. Gorden,et al.  Human growth hormone radioreceptor assay using cultured human lymphocytes. , 1973, Nature: New biology.

[2]  H. Thijssen.  A simple method for preparing 2-amino-4-hydroxy-6-formylpteridine, a precursor of the pteridine substrate of dihydropteroate biosynthesis. , 1973, Analytical biochemistry.

[3]  S. Lindskog,et al.  AMINO ACID COMPOSITION OF VARIOUS FORMS OF BOVINE AND HUMAN ERYTHROCYTE CARBONIC ANHYDRASE. , 1964, Biochimica et biophysica acta.

[4]  M. Horisberger.  Colloidal gold as a tool in molecular biology , 1983 .

[5]  E. F. Bowden,et al.  Electrochemical Aspects of Bioenergetics , 1985 .

[6]  W G Bardsley,et al.  The steady-state kinetics of peroxidase with 2,2'-azino-di-(3-ethyl-benzthiazoline-6-sulphonic acid) as chromogen. , 1975, The Biochemical journal.

[7]  L. Shannon,et al.  Peroxidase isozymes from horseradish roots. I. Isolation and physical properties. , 1966, The Journal of biological chemistry.

[8]  W. D. Geoghegan,et al.  Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[9]  V. Massey,et al.  Studies on milk xanthine oxidase. Some spectral and kinetic properties. , 1969, The Journal of biological chemistry.

[10]  J. Olson,et al.  The reaction of xanthine oxidase with lumazine. Characterization of the reductive half-reaction. , 1984, The Journal of biological chemistry.

[11]  V Massey,et al.  Tight binding inhibitors of xanthine oxidase. , 1981, Pharmacology & therapeutics.

[12]  T. Spector,et al.  Stoichiometric inhibition of reduced xanthine oxidase by hydroxypyrazolo [3,4-d]pyrimidines. , 1970, The Journal of biological chemistry.

[13]  G. Mabbott.  An introduction to cyclic voltammetry , 1983 .

[14]  A. L. Crumbliss,et al.  Preparation and activity of carbonic anhydrase immobilized on porous silica beads and graphite rods , 1988, Biotechnology and bioengineering.

[15]  G. Guilbault,et al.  Amperometric Determination of Hypoxanthine and Inosine by Use of Enzyme Sensors Based on a Clark-Type Hydrogen Peroxide or Oxygen Electrode , 1990 .

[16]  G. Holleck.  The reduction of chlorine on carbon in AlCl3-KCl-NaCl melts. , 1972 .

[17]  George G. Guilbault,et al.  Analytical Uses of Immobilized Enzymes , 1984 .

[18]  G. Frens.  Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions , 1973 .

[19]  N. Kjeldgaard,et al.  2-Amino-4hydroxy-6-formylpteridine, an inhibitor of purine and pterine oxidases. , 1950, Biochimica et biophysica acta.

[20]  V MASSEY,et al.  PURIFICATION AND PROPERTIES OF THE GLUCOSE OXIDASE FROM ASPERGILLUS NIGER. , 1965, The Journal of biological chemistry.

[21]  W. Heineman,et al.  Laboratory techniques in electroanalytical chemistry , 1984 .

[22]  M. Horisberger,et al.  Colloidal gold, a useful marker for transmission and scanning electron microscopy. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[23]  Y. Pocker,et al.  The catalytic versatility of erythrocyte carbonic anhydrase. 3. Kinetic studies of the enzyme-catalyzed hydrolysis of p-nitrophenyl acetate. , 1967, Biochemistry.

[24]  J H Luong,et al.  Development of a biosensor for assaying postmortem nucleotide degradation in fish tissues , 1990, Biotechnology and bioengineering.