Abstract
Fumonisin B(1) (FMB(1)) is a food-born mycotoxin produced by Fusarium moniliforme. Monoclonal antibody against FMB(1) (anti-FMB(1) mAb) was produced in the hybridoma DV9, which was established from a BALB/c mouse immunized with bovine serum albumin conjugated FMB(1) (FMB(1)-BSA). A competitive direct enzyme-linked immunosorbent assay (ELISA) showed that anti-FMB(1) mAb has about 10 ppb of minimum FMB(1) detection concentration and 220 ppb of 50% inhibition concentration (IC(50)). Much lower cross-reactivity of anti-FMB(1) mAb on ochratoxin A, aflatoxin B(1) and deoxynivalenol provided that anti-FMB(1) mAb was specific for FMB(1). The gene coding single chain variable fragment against FMB(1) (anti-FMB(1) scFv) was cloned from the hybridoma DV9 and was expressed in recombinant Escherichia coli. Insoluble anti-FMB(1) scFv required optimization of its refolding condition, and hence functional scFv was obtained. By using indirect ELISA, about 12-fold lower binding activity of anti-FMB(1) scFv on FMB(1)-BSA was obtained in comparison with that of the parental mAb.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aflatoxin B1 / chemistry
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Aflatoxin B1 / immunology
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology*
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Antibody Affinity / genetics
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Antibody Affinity / immunology
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Antibody Specificity / genetics
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Antibody Specificity / immunology*
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Cattle
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Cross Reactions / immunology
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Female
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Fumonisins / chemistry
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Fumonisins / immunology*
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Fusarium / chemistry
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Gene Expression
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Mice
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Mice, Inbred BALB C
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Ochratoxins / chemistry
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Ochratoxins / immunology
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Serum Albumin, Bovine / chemistry
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Serum Albumin, Bovine / immunology
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Single-Chain Antibodies / biosynthesis
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Single-Chain Antibodies / chemistry
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Single-Chain Antibodies / genetics
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Single-Chain Antibodies / immunology*
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Trichothecenes / chemistry
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Trichothecenes / immunology
Substances
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Antibodies, Monoclonal
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Fumonisins
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Ochratoxins
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Recombinant Proteins
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Single-Chain Antibodies
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Trichothecenes
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ochratoxin A
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Serum Albumin, Bovine
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fumonisin B1
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Aflatoxin B1
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deoxynivalenol