Preparation and identification of monoclonal antibody against fumonisin B(1) and development of detection by Ic-ELISA

Sumei Ling; Jie Pang; Jinjin Yu; Rongzhi Wang; Licai Liu; Yanling Ma; Yuming Zhang; Ni Jin; Shihua Wang

doi:10.1016/j.toxicon.2013.12.008

Preparation and identification of monoclonal antibody against fumonisin B(1) and development of detection by Ic-ELISA

Toxicon. 2014 Mar:80:64-72. doi: 10.1016/j.toxicon.2013.12.008. Epub 2013 Dec 27.

Authors

Sumei Ling¹, Jie Pang², Jinjin Yu², Rongzhi Wang¹, Licai Liu³, Yanling Ma¹, Yuming Zhang², Ni Jin¹, Shihua Wang⁴

Affiliations

¹ 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.
² School of Food Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
³ 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.
⁴ 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.

PMID: 24378835
DOI: 10.1016/j.toxicon.2013.12.008

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.

Keywords: Detection; Fumonisin B(1); Hybridoma; Monoclonal antibody; ic-ELISA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Monoclonal / biosynthesis*
Antibodies, Monoclonal / chemistry*
Cell Fusion
Cloning, Molecular
Enzyme-Linked Immunosorbent Assay / methods*
Female
Fumonisins / chemistry
Fumonisins / isolation & purification*
Hemocyanins
Inhibitory Concentration 50
Limit of Detection
Mice
Mice, Inbred BALB C

Substances

Antibodies, Monoclonal
Fumonisins
fumonisin B1
Hemocyanins
keyhole-limpet hemocyanin