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

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• 作者：Sumei Ling ; Jie Pang ; Jinjin Yu ; Rongzhi Wang ; Licai Liu ; Yanling Ma ; Yuming Zhang ; Ni Jin ; Shihua Wang
• 关键词：Fumonisin B1 ; Hybridoma ; Monoclonal antibody ; Detection ; ic-ELISA
• 刊名：Toxicon
• 出版年：15 March, 2014
• 年：2014
• 卷：80
• 期：Complete
• 页码：64-72
• 全文大小：1430 K

文摘

Fumonisin B₁ (FB₁) is one of the mycotoxins produced by Fusarium verticillioides, which was mainly found in corn and related products. FB₁ 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 FB₁-BSA was used to immunize Balb/c mice, and one hybrid cell line 4G5 excreting monoclonal antibody against FB₁ 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-FB₁ mcAb was harvested from ascites and the titer reached 6.4聽脳聽10⁴ after purification with caprylic/ammonium sulfate precipitation (CA-AS) method. The cross-reactivity results showed that anti-FB₁ mcAb was highly specific to fumonisin B₁, and the affinity was 2.1聽脳聽10⁸聽L/M. Indirect competitive ELISA (ic-ELISA) indicated that the linear range to detect FB₁ was 1-800聽ng/mL with IC50 of 32聽ng/mL. The detection limit was 1.0聽ng/mL, and the recovery average was 93.75聽卤聽6.90%. Therefore, the anti-FB₁ mcAb excreted by 4G5 can be used to detect fumonisin B₁ in corn and related samples.