Laccase from Coriolus hirsutus as alternate label for enzyme immunoassay

Determination of pesticide 2,4-dichlorophenoxyacetic acid
  • Anatoliy V. ZherdevEmail author
  • Nadezhda A. Bizova
  • Alexander I. Yaropolov
  • Natalija V. Lyubimova
  • Olga V. Morozava
  • Boris B. Dzantiev


A new label—laccase from the fungus Coriolus hirsutus—was applied for solid-phaseenzyme-linked immunosorbentassays of the pesticide 2,4-dichlorophenoxyacetic acid (2,4-D). Two proposed assays are based on (1) competitive binding of antibody-laccase conjugate with immobilized 2,4-D-protein conjugate and 2,4-D in tested sample, and (2) competition of 2,4-D and 2,4-D-laccase conjugate for binding with immobilized antibodies. Kinetic and concentration dependencies for these reactions were studied, and the ELISAs were optimized in accordance with the data obtained. The elaborated systems perm it the detection of 2,4-D in concentrations down to 10–20ng/mL; time of the assays is 1.5–2 h. The main advantage of the laccase label, in comparison with the widely used peroxidase one, lies in the lack of hydrogen peroxide from substrate mixture, because dissolved oxygen plays the role of oxidizer.

Index Entries

Enzyme immunoassay laccase pesticide 2,4-dichlorophen-oxyacetic acid 


  1. 1.
    Prince, C. P., and Newman, D. J. (1991), Principles and Practice of Immunoassay, Stockton, New York.Google Scholar
  2. 2.
    Vanderlaan, M., Stanker, L. H., Watkins, B. S., and Roberts, D. W. (1991), Immunoassays for Trace Chemical Analysis, American Chemical Society, Washington, DC.Google Scholar
  3. 3.
    Van Emon, J. M., and Lopez-Avila, V. (1992), Anal. Chem. 64, 79–89.CrossRefGoogle Scholar
  4. 4.
    Egorov, A. M., Reshetnikova, I. A., Fechina, V. A., and Gazaryan, I. G. (1995), Ann. NY Acad. Sci. 750, 469–472.CrossRefGoogle Scholar
  5. 5.
    Yaropolov, A. I, Skorobogat'ko, O. V., Vartanov, S. S., and Varfolomeyev, S. V. (1994), Appl. Biochem. Biotechnol. 49, 257–280.Google Scholar
  6. 6.
    Skorobogat'ko, O. V., Gindilis, A. L., Troitskaya, E N., Shuster, A. M., and Yaropolov, A. I. (1994), Anal. Lett. 27, 2997–3012.Google Scholar
  7. 7.
    Jung, F., Gee, S. J., Harrison, R. O., Goodrow, M. H., Karu, A. E., Braun, A. E., Li, Q. X., and Hammock, B. D. (1989), Pesticide Sci. 26, 303–317.CrossRefGoogle Scholar
  8. 8.
    Van Emon, J. M., and Mumma, R. O. (1990), ACS Symp. Ser. 442, 112–139.Google Scholar
  9. 9.
    Gazzaz, S. S., Rasco, B. A., and Dong, F. M. (1992), Crit. Rev. Food Sci. Nutr. 32, 197–229.CrossRefGoogle Scholar
  10. 10.
    Knopp, D. (1995), Anal. Chim. Acta. 311, 383–392.CrossRefGoogle Scholar
  11. 11.
    Gindilis, A. L., Zhazhina, E. O., Baranov, Yu. A., Kariakin, A. A., Gavrilova, V. P., and Yaropolov, A. I. (1988), Biochemistry (Moscow) 53, 635–639.Google Scholar
  12. 12.
    Dzantiev, B. B., Zherdev, A. V., Yulaev, M. F., Sitdikov, R. A., Dmitrieva, N. M., and Moreva, I. Yu. (1996), Biosensors Bioelectronics 11, 179–185.CrossRefGoogle Scholar
  13. 13.
    Habeeb A. F. S. (1996), Anal. Biochem. 14, 328–333.CrossRefGoogle Scholar
  14. 14.
    Fields, R. (1971), Biochem. J. 124, 581–590.Google Scholar
  15. 15.
    Ishikawa, E., Imagawa, M., Hashida, S., Yoshitaki, S., Hamaguchi, Y., and Ueno, T. (1983), J. Immunoassay 4, 209–327.CrossRefGoogle Scholar
  16. 16.
    Wilson, M. B., and Nakane, P. K. (1978), in Immunofluorescence and Related Staining Techniques, Knapp, W., Holubar, K., and Wick, G., eds., Elsevier/North-Holland Biochemical, Amsterdam, pp. 215–224.Google Scholar
  17. 17.
    Catty, D. (1989), Antibodies: A Practical Approach, IRL Press, Oxford, UK.Google Scholar
  18. 18.
    Rukavishnikova, G. E., Dzantiev, B. B., Liozner, A. L., Eremin, S. A., and Sigal, E. R. (1991), in Advances in Steroid Analysis '90, Gorog, S., ed., Akademiai Kiado, Budapest, pp. 103–108.Google Scholar
  19. 19.
    Dzantiev, B. B., Zherdev, A. V., Romanenko, O. G., Sapegova, L. A. (1996), Int. J. Environ. Anal. Chem. 65, 95–111.CrossRefGoogle Scholar
  20. 20.
    Van Oss, C. J., Good, R. J., and Chaudhyry, M. K. (1986), J. Chromatogr. 376, 111–119.CrossRefGoogle Scholar
  21. 21.
    Stevens, F. J. (1987), Mol. Immunol. 24, 1055–1060.CrossRefGoogle Scholar
  22. 22.
    Zherdev, A. V., Romanenko, O. G., and Dzantiev, B. B. (1997), J. Immunoassay, 18, 67–95.CrossRefGoogle Scholar
  23. 23.
    Fleeker, J. (1987), J. Assoc. Off. Anal. Chem. 70, 874–878.Google Scholar
  24. 24.
    Hall, J. C., Deschamps, R. J. A., and Krieg, K. K. (1989), J. Agric. Food Chem. 37, 1088–1093.CrossRefGoogle Scholar
  25. 25.
    Newsome, W. H. and Collins, P. G. (1989), Food Agric. Immunol. 1, 203–210.CrossRefGoogle Scholar
  26. 26.
    Franek, M., Kolar, V., Granatova, M., and Nevorankova, Z. (1994), J. Agric. Food Chem. 42, 1369–1374.CrossRefGoogle Scholar
  27. 27.
    Knopp, D. (1994), Occup. Environ. Med., 51, 153–159.Google Scholar
  28. 28.
    Meulenberg, E. P. and Stoks, P. G. (1995), Anal. Chim. Acta 311, 407–413.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc 1999

Authors and Affiliations

  • Anatoliy V. Zherdev
    • 1
    Email author
  • Nadezhda A. Bizova
    • 1
  • Alexander I. Yaropolov
    • 1
  • Natalija V. Lyubimova
    • 1
  • Olga V. Morozava
    • 1
  • Boris B. Dzantiev
    • 1
  1. 1.A. N. Bach Institute of BiochemistryRussian Academy of SciencesMoscowRussia

Personalised recommendations