Aflatoxin B1 (AFB1) is carcinogenic mycotoxin produced by Aspergillus flavus and Aspergillus parasiticus. Monoclonal antibody (mAb) specific to AFB1 were produced from a hybridoma cell line, 3F6, generated by the fusion of P3/NS1/1-AG4-1myeloma cells with spleen cells isolated from a BALB/c mouse immunized with AFB1-bovine serum albumin. The 3F6 mAb belongs to the immunoglobulin G1 isotype. A competitive direct enzyme-linked immunosorbent assay (cdELISA) and a competitive indirect ELISA (ciELISA) were established for antibody characterization. In the cdELISA, the concentrations causing 50% inhibition of binding of AFB1-horseradish peroxidase to the antibody by AFB1, AFB2, AFG1, and AFG2 were found to be 0.052, 0.050, 1.820, and 1.270 ng/mL, respectively. A rapid and sensitive mAb-based gold nanoparticle immunochromatographic strip (immunostrip) method was also established. This strip has a detection limit of 2.0 ng/ml for AFB1and can be completed within 10 min. Closely examining 16 samples by cdELISA revealed that 9 were slightly contaminated with AFB1 from 1.61~16.09 ng/g.  Results of 9 contaminated samples further analyzed with immunostrip assay correlate well with those obtained from cdELISA. The proposed cdELISAand immunostrip methods are highly sensitive to the rapid screening of AFB1 in food and feed samples.

Production of a monoclonal antibody against aflatoxin B1 and its application in enzyme-linked immunosorbent assay and rapid immunochromatographic strip

248th National Meeting (2014)

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