Production and characterization of antibody against diacetoxyscirpenol

Abstract

Antibodies against diacetoxyscirpenol (DAS) were obtained from rabbits after immunizing them with hemisuccinate or hemiglutarate derivatives of DAS conjugated to bovine serum albumin (BSA). DAShemiglutarate-BSA was found to be a much better immunogen than DAS-hemisuccinate-BSA. Competitive radioimmunoassay revealed that the antisera obtained from rabbits after immunization with DAS-hemiglutarate-BSA showed high specificity toward DAS. The concentrations causing 50% displacement of radioactive DAS by unlabeled DAS, 4-monoacetoxyscirpenol (MAS), and 15-MAS were found to be 1.5, 130, and 300 ng per assay, respectively. Thus, the cross-reactivities for 4-MAS and 15-MAS are ca. 87 and 300 times weaker than that of DAS. Practically no cross-reaction (less than 5% displacement) was observed when deoxynivalenol, T-2 toxin, deoxyverrucarol, and scirpentriol were tested at a concentration of 2,000 ng/ml.

Production and characterization of a monoclonal antibody cross-reactive with most group A trichothecenes

“…Detection of antibody titer in mouse serum-ascites fluids. The antibody titer of the mouse serum was determined by an RIA by using protocols essentially the same as those described before (3). Before the assay, the antiserum was precipitated with ammonium sulfate to a final saturation of 33.3%.…”

“…The approaches that have been generally used for the production of either polyclonal or monoclonal antibody against T-2 toxin involve the use of immunogens which were prepared by conjugation of T-2 toxin through the functional hydroxyl group at the C-3 position to a carrier protein (3,4,9,11,13,14,27). An alternative approach for conjugation of T-2 toxin to the carrier protein was made most recently in our laboratory (23).…”

Production and characterization of antibody against deoxyverrucarol

“…Tritiated sodium borohydride (77.9 Ci/mmol) was purchased from New England Nuclear Corp., Boston, Mass. Tritiated DAS was prepared in our laboratory (19.4 Ci/mmol) (2). The purity of these mycotoxins was further analyzed by a thin-layer chromatography (TLC) method (14) and was shown as a single spot at concentrations of 5 to 10 ,ug per spot.…”

“…However, because of the toxicity and the immunosuppressive effect of these mycotoxins, the antibody titers in rabbits were relatively low as compared with other haptens, such as steroids. In addition, the antibodies obtained from rabbits after being immunized with the respective mycotoxin-protein conjugate were very specific (2,3,5,14); thus, they cannot be used to determine other trichothecenes. To circumvent such problems, a recommendation was made by the Committee on Protection Against Mycotoxins of the National Research Council, urging use of deoxyverrucarol (DOVE)-protein conjugate as an immunogen for subsequent immunization (4).…”

Comparison of detection methods for trichothecenes produced by fusarium sporotrichioides on fodder and grains at different air humidities

“…Many strains of Fusan'urn sporotrichioides Sherb. have been reported to produce T-2 toxin and related trichothecenes, such as neosolaniol, IiT-2 toxin, and T-2 tetraol [Marasas et al, 1987;Bekele et al, 19911. Analytical methods based on thin layer chromatography (TLC), high performance liquid chromatography (HPLC), gas chromatography (GO, gas and liquid chromatography combined with mass spectrometry (GC-MS and LC-MS) [Scott, 1982;Rajakyll et al, 1987;Lauren and Agnew, 19911, and immunological techniques [Chu et al, 1984; 0 1996 Wtley-Ltss, Inc. Chiba et al, 1988;Usleber et al, 19911 are available for the determination of the four most frequently met trichothecenes: deoxynivalenol, T-2 toxin, diasetoxyscirpenol, and nivalenol.…”