Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Development of Homogeneous Enzyme Immunoassay for the Organophosphorus Insecticide Fenthion

  • Kim, Bok-Hee (Regional Innovation Center, Seowon University) ;
  • Park, Eun-Yong (Department of Biochemistry, Yeungnam University) ;
  • Lee, Yong-Tae (Department of Biochemistry, Yeungnam University) ;
  • Lee, Jung-Hun (Department of Biological Sciences, School of Biotechnology and Environmental Engineering, Myongji University) ;
  • Lee, Sang-Hee (Department of Biological Sciences, School of Biotechnology and Environmental Engineering, Myongji University)
  • Published : 2007.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

A rapid, convenient homogeneous competitive enzyme immunoassay for estimating the amount of fenthion is described. The assay utilizes glucose-6-phosphate dehydrogenase-hapten conjugates that are inhibited in solution by antibodies obtained from bovine serum albumin-hapten conjugates. In order to investigate the effects of bridging group recognition on the sensitivity of dose response characteristics, the bridging groups of varying alkyl chain length were attached at the phosphate position of fenthion. Among the antibodies used, the one obtained from the use of hapten (fenthion analog) with the same bridging group structure that was used in preparing the enzyme-fenthion conjugates showed maximum inhibition (up to 51.8%) in the absence of fenthion. In the presence of fenthion, the activity of the enzyme-hapten conjugate is regained in an amount proportional to the fenthion concentration. Under the optimized condition, the $ED_{50}$ value for fenthion was $0.809{\mu}g/ml$. The assay developed in this study is a rapid effective screening method for fenthion prior to precise analysis.

Keywords

References

  1. Abuknesha, R. A. and C. Luk. 2005. Paraquat enzymeimmunoassays in biological samples: Assessment of the effects of hapten-protein bridge structures on assay sensitivity. Analyst 130: 956-963 https://doi.org/10.1039/b418087a
  2. AOAC International. 1995. Official Methods of Analysis, Section 970.52, 18th Ed. Association of Official Analytical Chemists, Arlington, VA
  3. Bachas, L. G. and M. E. Meyerhoff. 1986. Theoretical models for predicting the effect of bridging group recognition and conjugate substitution on hapten enzyme immunoassay dose-response curves. Anal. Biochem. 156: 223-238 https://doi.org/10.1016/0003-2697(86)90177-6
  4. Brun, E. M., M. Garces-Garcia, R. Puchades, and A. Maquieira. 2004. Enzyme-linked immunosorbent assay for the organophosphorus insecticide fenthion: Influence of hapten structure. J. Immunol. Methods 295: 21-35 https://doi.org/10.1016/j.jim.2004.08.014
  5. Cho, Y. A., Y. J. Kim, B. D. Hammock, Y. T. Lee, and H. S. Lee. 2003. Development of a microtiter plate ELISA and a dipstick ELISA for the determination of the organophosphorus insecticide fenthion. J. Agric. Food Chem. 51: 7854-7860 https://doi.org/10.1021/jf0346915
  6. Eddleston, M., P. Eyer, F. Worek, F. Mohamed, L. Senarathna, L. von Meyer, E. Juszczak, A. Hittarage, S. Azhar, W. Dissanayake, M. H. Sheriff, L. Szinicz, A. H. Dawson, and N. A. Buckley. 2005. Differences between organophosphorus insecticides in human self-poisoning: A prospective cohort study. Lancet 366: 1452-1459 https://doi.org/10.1016/S0140-6736(05)67598-8
  7. Han, E.-M., H. R. Park, S. J. Hu, K.-S. Kwon, H. Lee, M.-S. Ha, K.-M. Kim, E.-J. Ko, S.-D. Ha, H. S. Chun, D.-H. Chung, and D.-H. Bae. 2006. Monitoring of aflatoxin B1 in livestock feeds using ELISA and HPLC. J. Microbiol. Biotechnol. 16: 643-646
  8. Hur, H. J., K. W. Lee, H. Y. Kim, D. K. Chung, and H. J. Lee. 2006. In vitro immunopotentiating activities of cellular fractions of lactic acid bacteria isolated from kimchi and bifidobacteria. J. Microbiol. Biotechnol. 16: 661-666
  9. Kim, B., G. S. Cha, and M. E. Meyerhoff. 1990. Homogeneous enzyme-linked binding assay for studying the interaction of lectins with carbohydrates and glycoproteins. Anal. Chem. 62: 2663-2668 https://doi.org/10.1021/ac00223a003
  10. Kim, B., I. Behbahani, and M. E. Meyerhoff. 1992. Lectinbased homogeneous enzyme-linked binding assay for estimating the type and relative amount of carbohydrate within intact glycoproteins. Anal. Biochem. 202: 166-171 https://doi.org/10.1016/0003-2697(92)90223-T
  11. Kim, B., J. M. Buckwalter, and M. E. Meyerhoff. 1994. Adapting homogeneous enzyme-linked competitive binding assays to microtiter plates. Anal. Biochem. 218: 14-19 https://doi.org/10.1006/abio.1994.1135
  12. Kim, K.-O., Y. J. Kim, Y. T. Lee, B. D. Hammock, and H. S. Lee. 2002. Development of an enzyme-linked immunosorbent assay for the organophosphorus insecticide bromophosethyl. J. Agric. Food Chem. 50: 6675-6682 https://doi.org/10.1021/jf025703+
  13. Kim, M. J., H. J. Kim, J. M. Kim, B. Kim, S. H. Han, and G. S. Cha. 1995. Homogeneous assays for riboflavin mediated by the interaction between enzyme-biotin and avidin-riboflavin conjugates. Anal. Biochem. 231: 400- 406 https://doi.org/10.1006/abio.1995.9980
  14. Kim, Y. J., Y. A. Cho, H.-S. Lee, Y. T. Lee, S. J. Gee, and B. D. Hammock. 2003. Synthesis of haptens for immunoassay of organophosphorus pesticides and effect of heterology in hapten spacer arm length on immunoassay sensitivity. Anal. Chim. Acta 475: 85-96 https://doi.org/10.1016/S0003-2670(02)01037-1
  15. Lee, D. H., B. J. Park, M. S. Lee, J. B. Choi, J. K. Kim, J. H. Park, and J.-C. Park. 2006. Synergistic effect of Staphylococcus aureus and LPS on silica-induced tumor necrosis factor production in macrophage cell line J774A.1. J. Microbiol. Biotechnol. 16: 136-140
  16. Lee, W., S.-S. Lim, B.-K. Choi, and J.-W. Choi. 2006. Protein array fabricated by microcontact printing for miniaturized immunoassay. J. Microbiol. Biotechnol. 16: 1216-1221
  17. Morozova, V. S., A. I. Levashova, and S. A. Eremin. 2005. Determination of pesticides by enzyme immunoassay. J. Anal. Chem. 60: 202-217 https://doi.org/10.1007/s10809-005-0075-0
  18. Paula, S., M. R. Tabet, C. D. Farr, A. B. Norman, and W. J. Jr. Ball. 2004. Three-dimensional quantitative structureactivity relationship modeling of cocaine binding by a novel human monoclonal antibody. J. Med. Chem. 47: 133-142 https://doi.org/10.1021/jm030351z
  19. Peruski, A. H. and L. F. Jr. Peruski. 2003. Immunological methods for detection and identification of infectious disease and biological warfare agents. Clin. Diagn. Lab. Immunol. 10: 506-513
  20. Petkam, R., R. Renaud, L. Lin, H. Boermans, and J. Leatherland. 2005. Effects of sub-lethal levels of dichlorodiphenyltrichloroethane and dichlorodiphenyldichloroethylene on in vitro steroid biosynthesis by ovarian follicles or steroid metabolism by embryos of rainbow trout (Oncorhynchus mykiss). Aquat. Toxicol. 73: 288-298 https://doi.org/10.1016/j.aquatox.2005.03.018
  21. Rubenstein, K. E., R. S. Schneider, and E. F. Ullman. 1972. 'Homogeneous' enzyme immunoassay. A new immunochemical technique. Biochem. Biophys. Res. Commun. 47: 846-851 https://doi.org/10.1016/0006-291X(72)90570-0
  22. Tetin, S. Y. and S. D. Stroupe. 2004. Antibodies in diagnostic applications. Curr. Pharm. Biotechnol. 5: 9-16 https://doi.org/10.2174/1389201043489602
  23. TomLin, C. D. S. 1997. The Pesticide Manual: A World Compendium, pp. 531-532, 11th Ed. The British Crop Protection Council, Surrey
  24. Wang, S., C. Yu, and J. Wang. 2005. Enzyme immunoassay for the determination of carbaryl residues in agricultural products. Food Addit. Contam. 22: 735-742 https://doi.org/10.1080/02652030500164326