Monoclonal Antibodies MAM-D3E4, MAM-D3C6, MAM-D12E2, and MAM-D4D6 Against Aflatoxins

摘要

Antigen Used for Immunization: Aflatoxins (AFs) are toxic secondary metabolites of the fungi from Aspergilus spp. Exposure to high levels of AFs may result in death, and long-term exposure to lower levels often leads to liver diseases, including liver cancer in humans. AFs are small molecular weight compounds that are not recognized by the mammalian immune system, that is, they are haptens. Immune response against these hapten structures can only be developed by conjugating them to a bigger and immunogenic molecule such as proteins.Conjugation of Aflatoxin B1 (AFB1) (Fermentek) to bovine serum albumin (BSA), ovalbumin (OVA), and human apotransferrin (TF) was achieved by modification of the method of Zhou et al. in two steps. In the first step, amine groups of the proteins were enriched by converting carboxyl groups to amine groups with an l-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) linker. In the second step, AFB1 was conjugated to the proteins by the Mannich-type reaction in the presence of formaldehyde. AFB1-TF and AFB1-0VA were used as immunogens and AFB1-BSA was used to test AF-specific immune response in indirect enzyme-linked immunosorbent assay (ELISA) test.