1996 Volume 1996 Issue 42 Pages 63-66
Novel monoclonal antibodies (mAb) against fumonisin B1 (FB1) were prepared and applied to an indirect competitive ELISA (ic-ELISA). BC6F1 female mice were immunized with a FB1-keyhole limpet hemocyanin conjugate (FB1-KLH). Trials of cell fusion yielded four hybridomas secreting mAb reactive with fumonisins (FB1-1, 2, 3, 4). FB1-2 mAb showing the highest reactivity to FB1 was used to construct an ic-ELISA. The detection limit was 0.2 ng FB1/ml in buffer solution. The relative cross-reactivities to FB2 and FB3 were 224% and 72% of FB1, respectively. Corn powder was firstly extracted with methanol-water (3 : 1), and the extract diluted with PBS (100 times) was directly applied to the ELISA. The recoveries of FB1 from corn spiked in range of 100-800 ppb FB1 were averaged to 88.4%. Corn samples (n=39) from China and Vietnam were analyzed for fumonisins by the present ic-ELISA, along with HPLC analysis, with a coefficient of correlation of 0.8. The detection limit of FB1 in this novel ELISA method combined with the one-step extraction procedure was 80 ppb, which is 10-60 fold lower than the ELISA previously repoted with anti-FB1 mAb.
