• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.111-117
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• 1991

We examined to develop the enzyme linked immunosorbent assay (ELISA) for determination of zearalenone in animal feeds. Zearalenone was first converted to 6'-(carboxymethyl) zearalenone oxime(zearalenone oxime) to get a coupling site and then conjugated to bovine serum albumin(BSA) for use as immunogen and to horseradish peroxidase(HRP) for use as enzyme marker. Antibody against zearalenone was obtained after 11 weeks of immunization of rabbit with zearalenone oxime-BSA. Cross reactivity of the antibody with ${\alpha}-zearalanol,\;{\beta}-zearalenol,\;{\alpha}-zearalanol\;and\;{\beta}-zearalanol$ were 168, 46, 26 and 20% respectiviely. A simple procedure was devised for the screening of zearalenone in feeds using ELISA. Feeds samples(5g) were extracted by blending with 25 ml of methanol-phospate butTered saline-dimethylformate(70 : 29 : 1) and the extract was filtered and aqueous filterate analyzed. It took only 1 hours to do whole procedure for the analysis of zearalenone in feeds by the direct competitive ELISA, and detectable limit was 1-100 ppb. Using this procedure, only 4 of 24 feed samples showed positive results with 3.93-7.43 ppb levels.

• Korean Journal of Food Science and Technology
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• v.41 no.1
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• pp.111-115
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• 2009

In this study we analysed for peanuts in processed foods using an enzyme-linked fluorescent immunoassay (ELFA), and compared the results with labeled ingredients. Crude peanut protein (CPP) was immunized into rabbits to produce specific antibodies(Ab). A sandwich ELFA was established using anti-CPP Ab and Ab-horseradish peroxidase (HRP) conjugate. The cross-reactivities of the Ab toward CPP, peanuts, almonds, soybeans, and walnuts were 100, 9.8, $1.1{\times}10^{-2},\;4.4{\times}10^{-3}$, and 0%, respectively. The samples included 19 items consisting of biscuits, snacks, chocolates, and so on. The results from the sandwich ELFA showed that peanuts were contained in 7 of the processed food items, among which, 5 items were labeled as having peanuts present but 2 items were not. One of the 2 items that was peanut-detected but unlabeled was a biscuit labeled to contain almonds and assayed to contain $2.1{\times}10^{-3}%$ peanuts, which might have been due to the weak cross-reactivity of the Ab toward almonds. The other item was a snack labeled to contain soybeans and assayed to contain 0.098% peanuts, which might have been due to peanut cross-contamination during processing, since the crossreactivity of the Ab toward soybeans was very weak. These results suggest that ELFA is a good tool to detect peanuts in processed foods, and allergens in certain processed foods should be labeled correctly.

• Korean Journal of Food Science and Technology
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• v.35 no.3
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• pp.366-372
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• 2003

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.