Production and Characterization of Monoclonal Antibodies Against Aflatoxin G 1

Abstract

Using semisolid methylcellulose HAT medium selection and non-competitive and competitive indirect enzyme-linked immunosorbent assay, three hybridomas that secreted antibodies for aflatoxin G₁ were selected after fusion of mouse SP2/0 myeloma cells with spleen cells isolated from BALB/c mice that had been immunized with the conjugate of aflatoxin G₁ (AFG₁) and bovine serum albumin. The aflatoxin-modified protein used to immunize mice was produced chemically by activating aflatoxin G₁ to a 9,10-epoxide derivative, which then covalently bound to the protein. All of these antibodies were found to be specific to AFG₁, and reacted strongly with aflatoxin G₂. These antibodies were designated 1C8, 1C10, and DE7, and had affinities for AFG1 of 5.1 × 10⁷, 2.7 × 10⁸, and 1.1 × 10⁸ liters mol⁻¹, respectively.