Browse > Article

Development of Enzyme-Linked Immunosorbent Assay for Glyphosate-Tolerant Soybeans  

Kwak, Bo-Yeon (Food Function Research Division, Korea Food Research Institute)
Ko, Seung-Hee (Food Function Research Division, Korea Food Research Institute)
Park, Chun-Wuk (Food Function Research Division, Korea Food Research Institute)
Son, Dae-Yeul (School of Medicine, Sungkyunkwan University)
Shon, Dong-Hwa (Food Function Research Division, Korea Food Research Institute)
Publication Information
Korean Journal of Food Science and Technology / v.35, no.3, 2003 , pp. 366-372 More about this Journal
Abstract
Enzyme-linked immunosorbent assay (ELISA) for assaying the 5-enolpyruvyshikimate-3-phosphate synthase from Agribacterium sp. CP4 (CP4 EPSPS) in genetically modified soybeans was developed. Polyclonal and monoclonal antibodies (Pab, Mab) specific to the CP4 EPSPS were produced. When using the Pab, the detection limit of sandwich ELISA toward CP4 EPSPS (0.03 ${\mu}g/mL$) was better than that of competitive indirect ELISA(ciELISA) (1 ${\mu}g/mL$). It was found that 2 of 3 monoclonal antibodies, Mab1 and Mab2, recognized the same antigenic determinant on CP4 EPSPS, but Mab3 recognized different antigenic determinant when competitive ELISA was performed using the Mabs. On the other hand, when the sensitivity of sandwich ELISA using combination of Pab and/or Mabs was determined, the sandiwich ELISA using Mab2 as a capture antibody and Pab-HRP as a secondary antibody showed the lowest detection limit of CP4 EPSPS (0.02 ${\mu}g/mL$). The sandwich ELISA developed in this study could be applied to detect glyphosate-tolerant soybeans.
Keywords
ELISA; glyphosate-tolerant soybeans; CP4 EPSPS; polyclonal antibody; monoclonal antibody;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Baird, D.D. Introduction of a new broad spectrum postemergence herbicide class with utility for herbaceous perennial weed control. North Central Weed Control Conf. 26: 64-68 (1971)
2 Steinruken, H.C. and Amrhein, N. The herbicide glyphosate is a potent inhibitor of 5-enolpyruvyl shikimic acid-3-phosphate synthase. Biochem. Biophys. Res. Commun. 94: 1207-1212 (1980)
3 Padgette, S.R., Taylor, N.B., Nida, D., Bailey, M.R., MacDonald, J., Holden, L. R. and Fuchs, R.L. Safety assessment of glyphosate- tolerant soybeans: The composition of the seeds is equivalentto conventional soybeans. J. Nutr. 126: 702-716 (1996)   PUBMED
4 Wurz, A., Bluth, A., Zeitz, P., Pfeifer, C. and Willmund, R. Quantitative analysis of genetically modified organisms (GMO) in processed food by PCR-based methods. Food Control 10: 385-389 (1999)   DOI   ScienceOn
5 Meyer, R. Development and application of DNA analytical methods for the detection of GMOs in foods. Food Control 10: 391-399 (1999)   DOI   ScienceOn
6 Coly, P.V., Wyatt, D., McCaughey, C. and O'Neill, H.J. A simple standardised protocol for the production of monoclonal antibodies against viral and bacterial antigens. J. Immunol. Meth. 153: 81-84 (1992)   DOI   ScienceOn
7 Kwak, B. Y., Kim, S. Y. and Shon, D. H. Detection of molds by enzyme-linked immunosorbent assay. J. Microbiol. Biotechnol. 9: 764-772 (1999)
8 Bradford, M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal. Biochem. 72: 248-254 (1976)   DOI   PUBMED   ScienceOn
9 Brett, G.M., Chambers, S.J., Huang, L. and Morgan, M.R.A. Design and development of immunoassays for detection of proteins. Food Control 10: 401-406 (1999)   DOI   ScienceOn
10 Barry, G., Kishore, G., Padgette, S., Taylor, M., Kolacz, K., Weldon, M., Re, D., Eichholtz, D., Fincher, K. and Hallas, L. Strategies for imparting glyphosate-tolerance to crop plants. pp. 139-145. In: Biosynthesis and Molecular Regulation of Amino Acids in Plants. Singh, B.K., Flores, H.E. and Shannon, J.C. (eds.). American Society of Plant Physiologists, Madison, Wisconsin USA (1992)
11 Haslam, E. Shikimic acid: Metabolism and Metabolites. p. 184. John Wiley & Sons, Chichester, England (1993)
12 Malik, J., Barry, G. and Kishore, G. The herbicide glyphosate. Biofactors 2: 17-25 (1989)   PUBMED
13 Padgette, S.R., Kolacz, K.H., Delannay, X., Re, D.B., Lavallee, B.I., Tinius, C.N., Rhodes, W.K., Otero, Y.I., Barry, G.F., Eichholtz, D.A., Peschke, Y.M., Nida, D.L., Taylor, N.B. and Kishore, G.M. Development, identification and characterization of a glyphosate- tolerant soybean line. Crop Sci. 35: 1451-1461 (1996)   DOI   ScienceOn
14 Rogan, G.J., Dudin, Y.A., Lee, T.C., Magin, K.M., Astwood, J.D., Bhakta, N.S., Leach, J.N., Sanders, P.R and Fuchs, R.L. Immunodiagnostic methods for detection of 5-enolpyruvylshikimate-3-phosphate synthase in Roundup Ready® sobybeans. Food Control 10: 407-414 (1999)   DOI   ScienceOn
15 Son, D.Y., Ahn, K.M. and Lee, S.I. Sequencing, cloning and expression of CP4EPSPS roundup ready soybean insert. Food Sci. Biotechnol. 12: 133-136 (2003)