A 1-step enzyme-linked Immunosorbent assay (EUSA) method, using a highly sensitive and specific monoclonal antibody to aflatoxin B1 (AFB1), was evaluated by comparison with other methods, Including liquid chromatography (LC) and thinlayer chromatography. The detection limit of the ELISA was as low as 100 pg/assay. AFB1 contents of naturally contaminated corn samples were determined by the 3 methods. The relationships among the methods were Investigated, and good correlations were observed. Mixed feeds were also subjected to AFB1 determination by the 3 methods. For our ELISA system, 3 types of sample preparations were tested. For analysis of mixed feed by ELISA, samples must first be purified by column chromatography. When the relationship between LC and ELISA was also investigated, results were found to have a good correlation coefficient.
A direct enzyme-linked immunosorbent assay (ELISA) for the quantitation of aflatoxin B1 (AFB1) was developed by the utilization of horseradish peroxidase (HRP) labelled anti-AFB1 monoclonal antibody and 96-well immunomicroplate coated with AFB1-bovine serum albumin conjugate. The detection limit for AFB1 was ca. 5 pg/assay.A quick ELISA system was also developed using the 8-well immunomicroplate, and ca.50 pg to 5 ng/assay of AFB1 were quantitated within 10 min.Aflatoxin B1 (AFB1) is the well-known toxic metabolite of Aspergillus flavus and A.parasiticus, and exhibits an extensive toxicity and carcinogenicity. For the analysis of AFB1, thin-layer chromatography (TLC) was first developed by the utilization of the fluorescence of AFB1 20 years ago. This method was adopted as an official analysis, and currently, high performance liquid chromatography (HPLC) is introduced, since the sensitivity and resolution of HPLC are much higher than those of TLC. Chemical clean-up steps after the extraction of samples were essential for the accurate analysis of AFB, in both TLC and HPLC. As clean-up steps take a lot of time for the analysis, some immunoassays using antibody against AFB1 have been studied so far. Radioimmunoassay using polyclonal antibody was performed by Chu et al. in 1981. They reported that the sensitivity of AFB1 was in the range of 30-50 pg/assay.Monoclonal antibody against AFBI was used in the radioimmunoassay by Groopman et al. and 1 ng of AFB1 was analysed. Ueno developed an enzyme-linked immunosorbent assay (ELISA) using polyclonal antibody and reported the sensitivity of 10 pg/assay.We have already reported the production of monoclonal antibodies and the establishment of ELISA for ochratoxin A and T-2 toxin. In this paper, we report the ELISA system for the quantitation of AFBI using horseradish peroxidase (HRP) labelled anti-AFB1 mono-
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