Preparation of aflatoxin B1-BSA conjugate with high hapten/carrier molar ratio

We reported previously on the properties of an anti-aflatoxin B1 (AFB) antiserum raised with a conjugate in which AFB is coupled at the C8 position to bovine serum albumin (BSA). The hapten/carrier protein molar ratio of the conjugate was 14. We have since been able to optimize the coupling conditions, resulting in the preparation of another conjugate, with a molar ratio of 45. Comparison of the 2 conjugates by double-gel immunodiffusion analysis shows that little BSA 'activity' remains on the conjugate with a high molar ratio; on the other hand, its capacity to precipitate in the presence of anti-AFB antibodies is increased.