Polyclonal and monoclonal antibodies against fusarenon‐X were prepared by using a fusarenon‐X oxime derivative coupled to human serum albumin as the antigen for the immunization of rabbits and BALB/c mice, respectively. The specificity and sensitivity of these antibodies were tested by using fusarenon‐X oxime coupled to horseradish peroxidase as an enzyme‐linked toxin in a competitive assay with a double antibody solid phase. The relative cross‐reactivities of the polyclonal antiserum with T‐2 toxin, diacetoxyscirpenol, fusarenon‐X and neosolaniol were 3.67, 2.75, 1.0 and 0.58, respectively. The monoclonal antibody, however, showed considerable cross‐reactivity only with neosolaniol (0.28). The detection limit for fusarenon‐X was 150 pg/ml (5 pg per assay) and 300 pg/ml (10 pg per assay), respectively, using the polyclonal and monoclonal antibodies.

Thirteen monoclonal antibodies reactive with HT-2 were prepared by using a HT-2 hemisuccinate coupled to human serum albumin as antigen for the immunization of BALB/c mice. In a competitive enzyme immunoassay on a double antibody solid phase using HT-2 hemisuccinate coupled to horseradish peroxidase as enzyme linked toxin all antibodies reacted much better with T-2 toxin and acetyl T-2 than with HT-2. Eleven antibodies showed almost the same sensitivity and specificity, and one of these, designated 3E2, is extensively described. Its cross-reactivities with HT-2, T-2 toxin, acetyl T-2, iso T-2, T-2 tetraol tetraacetate and T-2 triol were 1·0, 140·2, 161·2, 0·32, 0·14 and 0·016, respectively. Two other antibodies, designated 2A4 and 2A5, behaved quite differently. The cross-reactivities of antibody 2A4 with these toxins were: 1·0, 113·9, 374·4, 1·35, 0·34 and 0·023, respectively; for antibody 2A5 they were 1·0, 46·1, 155·4, 8·31, 0·9 and 0·08, respectively. All antibodies proved to be IgGl. By using the antibody 3E2 a highly sensitive and very specific enzymc immunoassay for the detection of T-2 toxin was developed. The detection limit for T-2 toxin was 5 pg/ml (0·25 pg/assay).

A monoclonal antibody to the trichothecene mycotoxin diacetoxyscirpenol (DAS) was produced by a hybridoma, designated 2E5. It secreted antibody of the IgGl subclass and had a detection limit for DAS of 16 ng/ml with a direct enzyme immunoassay on a double antibody solid phase. The relative cross‐reactivities with 3α‐acetyl‐DAS, diacetylverrucarol, neosolaniol, T‐2 tetraol tetraacetate, fusarenon X, T‐2 toxin, and HT‐2 were 2224·5, 53·7, 13·9, 9·2, 6·4, 1·7, 0·6, and 0·35%, respectively.

Two murine monoclonal antibodies to the macrocyclic trichothecene roridin A are described. Screening for antibody production was performed on absorbed anti-mouse immunoglobulin serum as double-antibody solid phase, and further characterization was done on affinity-purified anti-mouse IgG serum. The antibodies, designated 5G11 and 4H10, had affinity constants for roridin A of 9.25 X 10(7) and 1.7 X 10(7) liters/mol, respectively. In monoclonal antibody-based direct enzyme immunoassays, these IgG1 antibodies had detection limits for roridin A of 0.4 ng/ml (0.02 ng per assay) and 1.8 ng/ml (0.09 ng per assay), respectively. Both antibodies were most specific for the tested macrocyclic trichothecenes. The relative cross-reactivities of antibody 5G11 with roridin A, roridin J, verrucarin A, satratoxin G, and satratoxin H were 100.0, 43.8, 16.7, 3.7, and 18.9%, respectively; for antibody 4H10 they were 100.0, 6.3, 64.0, 4.4, and 4.9%, respectively.

SummaryA new method for the screening of monoclonal antibodies to mycotoxins was developed using a double antibody solid phase in a direct enzyme immunoassay. Wells of microtitre plates were coated with affinity‐purified anti‐mouse IgG antiserum. The monoclonal antibody against the trichothecene T‐2 toxin bound to this solid phase was detected by reaction with an HT‐2 toxin‐peroxidase conjugate. The monoclonal antibody belongs to the IgG 1 subclass and has a detection limit for T‐2 toxin of 50ng/ml in a competitive direct enzyme immunoassay. The relative cross‐reactivity with acetyl T‐2, HT‐2, iso T‐2, T‐2 triol and T‐2 tetraol‐tetraacetate was 0.75, 0.35, 0.35, 0.22 and 0.01, respectively. No cross‐reactions with other trichothecenes could be found.ZusammenfassungEin monoklonaler Antikörper gegen das Trichothecen T‐2 Toxin: Screening des Antikörpers mit einem direkten EnzymimmunoassayFür das Screening von monoklonalen Antikörpern gegen Mykotoxine wurde eine neue Methode entwickelt.Es wird ein direkter Enzymimmunoassay mit einer “Double antibody solid phase” verwendet, bei dem die Kavitäten der Mikrotiterplatte mit affinitätschromatographisch gereinigtem Anti‐Maus IgG‐Antiserum beschichtet werden. Der gebundene monoklonale Antikörper gegen T‐2 Toxin wurde durch die Reaktion mit einem HT‐2 Toxin‐Peroxidase‐Konjugat nachgewiesen.Der monoklonale Antikörper gehörte zur IgG 1‐Subklasse und hatte im direkten kompetitiven Enzymimmunoassay eine Nachweisgrenze von 50 ng/ml für T‐2 Toxin. Die relative Kreuzreaktivität mit Acetyl T‐2, HT‐2, Iso T‐2, T‐2 Triol und T‐2 Tetraoltetraacetat betrug 0,75, 0,35, 0,35, 0,22 und 0,01. Mit anderen Trichothecenen wurden keine Kreuzreaktionen gefunden.