Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Development of Enzyme-Linked Immunosorbent Assay for the Detection of Fumonisins

효소면역측정법을 이용한 Fumonisin의 검출법 개발

  • Shon, Dong-Hwa (Food Chemistry and Physics Division, Korea Food Research Institute) ;
  • Hahn, Seong-Min (Research Center for New Bio-Materialsi in Agriculture, College of Agriculture and Life Sciences, Seoul National University) ;
  • Lim, Sun-Hee (Research Center for New Bio-Materialsi in Agriculture, College of Agriculture and Life Sciences, Seoul National University) ;
  • Lee, Yin-Won (Research Center for New Bio-Materialsi in Agriculture, College of Agriculture and Life Sciences, Seoul National University) ;
  • Cho, Sun-Hee (Department of Vertenary, Chungbuk National University) ;
  • Kang, Shin-Young (Department of Vertenary, Chungbuk National University) ;
  • Lee, Kyung-Ae (Food Chemistry and Physics Division, Korea Food Research Institute)
  • 손동화 (한국식품개발원, 이화학연구부) ;
  • 한성민 (농업생물신소재연구센터, 서울대학교 농업생명과학대학) ;
  • 임선희 (농업생물신소재연구센터, 서울대학교 농업생명과학대학) ;
  • 이인원 (농업생물신소재연구센터, 서울대학교 농업생명과학대학) ;
  • 조선희 (충북대학교 수의학과) ;
  • 강신영 (충북대학교 수의학과) ;
  • 이경애 (한국식품개발원, 이화학연구부)
  • Published : 1996.02.01
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Abstract

In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

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