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

제초제내성 유전자재조합 콩의 검출을 위한 면역분석법 개발

  • 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)
  • 곽보연 (한국식품개발연구원 식품기능연구본부) ;
  • 고승희 (한국식품개발연구원 식품기능연구본부) ;
  • 박춘욱 (한국식품개발연구원 식품기능연구본부) ;
  • 손대열 (성균관대학교 의과대학) ;
  • 손동화 (한국식품개발연구원 식품기능연구본부)
  • Published : 2003.06.01

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

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