title: Preparation and identification of monoclonal antibody against fumonisin B(1) and development of detection by Ic-ELISA.
abstract: 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.
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Sumei
authors.initials: S
authors.lastname: Ling
authors.affiliation: School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Pang
authors.affiliation: School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Jinjin
authors.initials: J
authors.lastname: Yu
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Rongzhi
authors.initials: R
authors.lastname: Wang
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China; Environmental Protection Bureau of Qitaihe City, Qitaihe 154600, Heilongjiang, China.
authors.forename: Licai
authors.initials: L
authors.lastname: Liu
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Ma
authors.affiliation: School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Yuming
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
authors.forename: Ni
authors.initials: N
authors.lastname: Jin
authors.affiliation: Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China. Electronic address: wshyyl@sina.com.
authors.forename: Shihua
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry*; D002459:Cell Fusion; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D037341:Fumonisins / Q000737:chemistry / Q000302:isolation & purification*; D006433:Hemocyanins; D020128:Inhibitory Concentration 50; D057230:Limit of Detection; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D037341:Fumonisins; C056933:fumonisin B1; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin
keywords: Detection; Fumonisin B(1); Hybridoma; Monoclonal antibody; ic-ELISA