Studies on the Immunochemical Techniques for Detection of Selected Fungal and Dinoflagellate Toxins
Annual progress rept. no. 2, 1 Aug 1982-31 Jul 1983
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During the second year August 1, 1982 to July 31, 1983, conditions for improving antibody production against T-2 toxin were studied. T-2 HS-BSA which has 10-15 moles of T-2 per mole of BSA was found to be a better immunogen than T-2-HG-BSA. Antibody titers as high as 11,000-14,000 were obtained from rabbits after repeated booster injections. High specific radioactive T-2, DAS, DOVE were prepared. With combination of the high specific radioactive T-2 toxin and high titer antibody, as little as 25 pg of T-2 toxin can be detected by the RIA in each assay. An indirect ELISA which permits detection of 0.2-1 ppb of T-2 toxin in urine, serum and milk was also established. Antibody against T-2 toxin has been used to localize T-2 toxin in different tissues and organs of mice by an immunohistochemical technique developed in our laboratory. A collaborative study with Dr. Hunter revealed that antibody titers of T-2 toxin increased drastically in an immunomodulation system in mice. Titers as high as 50,000 were obtained in mice 10 days after immunization with T-2-HS-IgG goat. Antibodies against diacetoxyscirpenol DAS, and deoxyverrucarol DOVE were obtained from rabbits after immunizing with DAS-hemisuccinate HS-BSA, DAS-hemiglutarate HG-BSA and DOVE-HS-BSA. DAS-HG-BSA was found to be a better immunogen than DAS-HS- BSA for the production of antibody against DAS. The antibody for DAS is most specific for DAS. However, DOVE antidbodies were found to be less specific. Radioimmunoassays for both DAS and DOVE were established, and the detection limits were found.
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