Sensitive immunoassay for fumonisin B1 in corn
1996
Yeung, J.M. | Prelusky, D.B. | Savard, M.E. | Dang, B.D.M. | Robinson, L.A.
Polyclonal antibodies against fumonisin B1 (FB1) were produced in rabbits after immunization with FB1 conjugates. Protection of the amino group prior to coupling the carboxylic acid of FB1 to human serum albumin, and then deprotecting the amine before immunization, generated high-affinity antibodies. The antibodies produced were specific to fumonisins and their hydrolyzed products (HFB1, HFB2) but not to any other mycotoxins commonly found in grains. The relative cross-reactivities of FB1, fumonisin B2 (FB2), fumonisin B3 (FB3), HFB1, and HFB2 were 100%, 55%, 13%, 11%, and 3%, respectively. Using these antibodies, a sensitive indirect competitive ELISA was developed. The detection limit and IC50 for FB1 in water were 0.05 and 0.66 ng/mL, respectively. The linear range was determined to be 0.05-0.75 ng/mL. The method was applied directly to 75% methanol extracts of corn. Recoveries of FBI were better than 87% from 10 to 10 000 ng/g at five levels of fortification. The detection limit in corn was 5 ng/g.
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