Production of ultrasensitive antibodies against aflatoxin B 1

Abstract

Aims: To produce speciÆc antibodies against the haptenic fungal toxin aØatoxin B1 (AFB1) and apply these antibodies in immunochemical assays for aØatoxins. Methods and Results: Rabbits were immunized using an AFB1 -bovine serum albumin conjugate and serum titres determined by double-antibody enzyme immunoassay. High titres of antibodies with very high afÆnity for AFB 1 were obtained 15 and 4 weeks after the initial immunization and the Ærst booster immunization respectively. The antibodies were employed in enzyme immunoassay (EIA) and immunoafÆnity chromatography (IAC) methods for aflatoxins. With a detection limit of 15 · 8pgml)1 for AFB1 , the EIA employing these antibodies is the most sensitive test for AFB1 described so far. In IAC columns, these antibodies provided high binding capacity for all major aØatoxins, including AFB1 , AFB2 , AFG1 and AFG2

Conclusions: The antibodies described here are useful for the analysis of trace levels of aflatoxins. SigniÆcance 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.