Production of ultrasensitive antibodies against aflatoxin B-1

Aims: To produce specific antibodies against the haptenic fungal toxin afla toxin B-1 (AFB(I)) and apply these antibodies in immunochemical assays for aflatoxins. Methods and Results: Rabbits were immunized using an AFB(1)-bovine serum al bumin conjugate and serum titres determined by double-antibody enzyme immun oassay. High titres of antibodies with very high affinity for AFB(1) were o btained 15 and 4 weeks after the initial immunization and the first booster immunization respectively. The antibodies were employed in enzyme immunoas say (EIA) and immunoaffinity chromatography (IAC) methods for aflatoxins. W ith a detection limit of 15.8 pg ml(-1) for AFB(1), the EIA employing these antibodies is the most sensitive test for AFB(1) described so far. In IAC columns, these antibodies provided high binding capacity for all major afla toxins, including AFB(1), AFB(2), AFG(1) and AFG(2). Conclusions: The antibodies described here are useful for the analysis of t race levels of aflatoxins. Significance and Impact of the Study: Polyclonal antibody-based EIA and IAC methods for aflatoxin analysis offer a suitable alternative to the more ex pensive monoclonal antibody-based methods.