Preparation and identification of monoclonal antibody against fumonisin B1 and development of detection by Ic-ELISA

• DOI: 10.1016/j.toxicon.2013.12.008
• Publication Date: 2014-01-02T23:35:47Z
• Authors: Sumei Ling; Jie Pang; Jinjin Yu; Rongzhi Wang; Licai Liu; Yanling Ma; Yuming Zhang; Ni Jin; Shihua Wang
• Keywords: Mice, Inbred BALB C; Antibodies, Monoclonal; Enzyme-Linked Immunosorbent Assay; 04 agricultural and veterinary sciences; Fumonisins; 3. Good health; Cell Fusion; Inhibitory Concentration 50; Mice; Limit of Detection; Hemocyanins; Animals; Female; Cloning, Molecular; 0405 other agricultural sciences

Fumonisin B(1) (FB(1)) is one of the mycotoxins produced by Fusarium verticillioides, which was mainly found in corn and related products. FB(1) was small molecule with no immunogenicity, so it should be conjugated to carrier proteins such as BSA (bovine serum albumin) or KLH (keyhole limpet hemocyanin) to generate immunogenicity. In this study, conjugate FB(1)-BSA was used to immunize Balb/c mice, and one hybrid cell line 4G5 excreting monoclonal antibody against FB(1) was obtained by fusing mouse Sp2/0 myeloma cells with spleen cells from the immunized mouse. Hybridoma 4G5 was injected into the abdomen of Balb/c mice, and the anti-FB(1) mcAb was harvested from ascites and the titer reached 6.4 × 10(4) after purification with caprylic/ammonium sulfate precipitation (CA-AS) method. The cross-reactivity results showed that anti-FB(1) mcAb was highly specific to fumonisin B(1), and the affinity was 2.1 × 10(8) L/M. Indirect competitive ELISA (ic-ELISA) indicated that the linear range to detect FB(1) was 1-800 ng/mL with IC50 of 32 ng/mL. The detection limit was 1.0 ng/mL, and the recovery average was 93.75 ± 6.90%. Therefore, the anti-FB(1) mcAb excreted by 4G5 can be used to detect fumonisin B(1) in corn and related samples.