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Applied Biochemistry and Biotechnology

, Volume 74, Issue 1, pp 1–12 | Cite as

Covalently immobilized choline oxidase and cholinesterases on a methacrylate copolymer for disposable membrane biosensors

  • Lucio Doretti
  • Paola Gattolin
  • Anna Burla
  • Daniela Ferrara
  • Silvano Lora
  • Giancarlo Palma
Original Articles

Abstract

Bienzymatic sensors for the determination of esters of choline were prepared by covalent co-immobilization of cholinesterases and choline oxidase on polymer membranes, obtained by radiation-induced copolymerization of 2-hydroxyethyl methacrylate and glycidyl methacrylate at low temperature. Optimization of the covalent attachment of choline oxidase and acetyl-or butyrylcholinesterase to copolymer was explored. The enzyme-modified polymers were applied on platinum electrodes to form amperometric sensors, based on the electrochemical detection of enzymatically developed hydrogen peroxide. Acetyl-, acetylthiobutyryl-, and butyrylthiocholine contents in standard solutions were measured, and linear calibration curves were determined. Temperature and pH effects on the electrochemical response are described.

Index entries

Biosensors acetylcholinesterase butyrylcholinesterase choline oxidase covalent immobilization radiation induced polymerization 

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References

  1. 1.
    Yao, T. (1983),Anal. Chim. Acta 153, 169–174.CrossRefGoogle Scholar
  2. 2.
    Mascini, M. and Moscone, D. (1986),Anal. Chim. Acta 179, 439–444.CrossRefGoogle Scholar
  3. 3.
    Tamiya, E., Sugiura, Y., Navera, E. N., Miroshita, S., Nakajima, K., Akiyama, A., and Karube, I. (1991),Anal. Chim. Acta 251, 129–134.CrossRefGoogle Scholar
  4. 4.
    Rouillon, R., Mionetto, N., and Marty, J. L. (1992),Anal. Chim. Acta 268, 347–350.CrossRefGoogle Scholar
  5. 5.
    Marty, J. L., Sode, K., and Karube, I. (1989),Anal. Chim. Acta 228, 49–53.CrossRefGoogle Scholar
  6. 6.
    Lopez, Ruiz, B., Dempsey, E., Hua, C., Smyth, M. R., and Wang, J. (1993),Anal. Chim. Acta 273, 425–430.CrossRefGoogle Scholar
  7. 7.
    Huang, Z., Villarta-Snow, R., Lubrano, G. J., and Guibault, G. G. (1993),Anal. Biochem. 215, 31–37.CrossRefGoogle Scholar
  8. 8.
    Navera, E. N., Suzuki, M., Tamiya, E., Takeuchi, T., and Karube, I. (1993),Electroanalysis 5, 17–22.CrossRefGoogle Scholar
  9. 9.
    Kano, K., Morikage, G., Uno, B., Esaka, Y., and Goto, M. (1994),Anal. Chim. Acta 299, 69–76.CrossRefGoogle Scholar
  10. 10.
    Morelis, R. M. and Coulet, P. R. (1990),Anal. Chim. Acta 231, 27–32.CrossRefGoogle Scholar
  11. 11.
    Skladal, P. and Mascini, M. (1992),Biosensors Bioelectron. 7, 335–343.CrossRefGoogle Scholar
  12. 12.
    Bernabei, M., Chiavarini, S., Cremisini, C., and Palleschi, G. (1993),Biosensors Bioeledron. 8, 265–271.CrossRefGoogle Scholar
  13. 13.
    Imato, T. and Ishibashi, N. (1995),Biosensors Bioelectron. 10, 435–441.CrossRefGoogle Scholar
  14. 14.
    Kumaran, S. and Morita, M. (1995),Talanta 42, 649–655.CrossRefGoogle Scholar
  15. 15.
    Cremisini, C., Di Sario, S., Mela, J., Pilloton, R., Palleschi, G. (1995),Anal. Chim. Acta 311, 273–280.CrossRefGoogle Scholar
  16. 16.
    Trojanowicz, M. and Hitchman, M. L. (1996),Trends Anal. Chem. 15, 38–45.Google Scholar
  17. 17.
    Palchetti, I., Cagnini, A., Del Carlo, M., Coppi, C., Mascini, M., and Turner A. P. F. (1997),Anal. Chim. Acta 337, 315–321.CrossRefGoogle Scholar
  18. 18.
    Doretti, L., Ferrara, D., Barison, G., and Lora, S. (1994),Appl. Biochem. Biotech. 49, 191–202.CrossRefGoogle Scholar
  19. 19.
    Doretti, L., Gattolin, P., and Lora, S. (1994),Anal. Lett. 27, 2455–2470.Google Scholar
  20. 20.
    Sundberg, L. and Porath, J. (1974),J. Chromat. 90, 87–98.CrossRefGoogle Scholar
  21. 21.
    Kalal, J. (1978),J. Polym. Sci. Polym. Symp. 62, 251–255.Google Scholar
  22. 22.
    Hannibal-Friedrich, O., Chun, H., and Sernetz, M. (1980),Biotechnol. Bioeng. 22, 157–175.CrossRefGoogle Scholar
  23. 23.
    Svec, F., Kalal, J., Menyailova, I.I., and Nakhapetyan, L. A. (1978),Biotechnol. Bioeng. 20, 1319–1328.CrossRefGoogle Scholar
  24. 24.
    Hall, C. E. and Hall, E. A. H. (1993),Anal. Chim. Acta 281, 645–653.CrossRefGoogle Scholar
  25. 25.
    Hall, C. E. and Hall, E. A. H. (1995),Anal. Chim. Acta 310, 199–210.CrossRefGoogle Scholar
  26. 26.
    Hall, C. E., Datta D., and Hall, E. A. H. (1996),Anal. Chim. Acta 323, 87–96.CrossRefGoogle Scholar
  27. 27.
    Doretti, L., Ferrara, D., Gattolin, P., and Lora, S. (1996),Biosensors Bioelectron. 11, 365–373.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc 1998

Authors and Affiliations

  • Lucio Doretti
    • 1
  • Paola Gattolin
    • 1
  • Anna Burla
    • 1
  • Daniela Ferrara
    • 1
  • Silvano Lora
    • 2
  • Giancarlo Palma
    • 3
  1. 1.Ufficio Sicurezza e Prevenzione del CNRPadovaItaly
  2. 2.Sezione di LegnaroIstituto di Fotochimica e Radiazioni d’Alto Energia del CNRLegnaro (Padova)Italy
  3. 3.Dipartimento di Chimica FisicaUniversità di PodovaPadovaItaly

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