Production of ultrasensitive antibodies against aflatoxin B 1

Aims: To produce specific antibodies against the haptenic fungal toxin aflatoxin B 1 (AFB 1 ) and apply these antibodies in immunochemical assays for aflatoxins. Methods and Results: Rabbits were immunized using an AFB 1 -bovine serum albumin conjugate and serum titres determined by double-antibody enzyme immunoassay. High titres of antibodies with very high affinity for AFB 1 were obtained 15 and 4 weeks after the initial immunization and the first booster immunization respectively. The antibodies were employed in enzyme immunoassay (EIA) and immunoaffinity chromatography (IAC) methods for aflatoxins. With 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 aflatoxins, including AFB 1 , AFB 2 , AFG 1 and AFG 2 . Conclusions: The antibodies described here are useful for the analysis of trace 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 expensive monoclonal antibody-based methods.