Immunochemical Methods for Fumonisins

Chu, Fun Sun

doi:10.1007/978-1-4899-1379-1_11

Cited by 6 publications

(2 citation statements)

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“…The availability of specific and sensitive mAb provides an unlimited supply of immunochemical reagents for enzyme-linked immunosorbent assay (ELISA) and allows further investigation of antibody structure. However, most of the mAbs developed appear to have a low affinity to fumonisins (Chu, 1996). Thus, the sensitivity of most mAb-based immunoassays for fumonisins was low, with a detection limit in the range of 0.1-1 ppm.…”

Section: Introductionmentioning

confidence: 99%

Production and Characterization of Monoclonal Antibodies against Fumonisin B1

Yu¹,

Chu²

1999

Food and Agricultural Immunology

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Monoclonal antibodies (mAb) against fumonisin B1 (FmB1) were produced from a stable hybridoma cell line, 9C11E6, generated by the fusion of P3/NS-1/1-AG4-1 myeloma cells with spleen cells isolated from a BALB/c mouse immunized with FmB1-keyhole limpet hemocyanin. The 9C11E6 mAb belong to the immunoglobulin G1 (kappa chain) isotype with the highest specificity for FmB3. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were established for antibody characterization and toxin analysis. The concentrations causing 50% inhibition (IC 50) of binding of FmB1horseradish peroxidase with the antibodies by FmB1, FmB2, and FmB3 in the dc-ELISA were found to be 29.1, 17.5, and 4.1 ng ml-1 , respectively. The IC 50 values of the binding of 9C11E6 mAb to the solid-phase FmB1-ovalbumin by free FmB1, FmB2, and FmB3 in the idc-ELISA were found to be 20.1, 13.2, and 16.8 ng ml-1 , respectively. The average recovery of FmB1 (50-2000 ng g-1) added to cornmeal and subsequently extracted with phosphatebuffered saline in the dc-ELISA was found to be 71.3%. The mAb did not react with the hydrolyzed FmB1, sphingosine, sphinganine, or tricarballylic acid, but showed weak crossreactivity (about 6.8% of FmB1) with the Alternaria alternata f. sp. lycopersici toxin TA.

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Section: Introductionmentioning

confidence: 99%

Production and Characterization of Monoclonal Antibodies against Fumonisin B1

Yu¹,

Chu²

1999

Food and Agricultural Immunology

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“…The most sensitive assays, with limits of detection in the low ppb range, have relied upon PAbs rather than MAbs (Usleber et al, 1994;Yu & Chu, 1996). An excellent review of current immunochemical methods for fumonisins was published recently (Chu, 1996). The commercial ELISAs typically take 30 min to several hours to complete.…”

Section: Introductionmentioning

confidence: 99%

Detection of the mycotoxin fumonisin B₁by a combination of immunofluorescence and capillary electrophoresis

Maragos

1997

Food and Agricultural Immunology

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The specificity of antibodies has been combined with the speed and resolving power of capillary electrophoresis for application to the analysis of the mycotoxin fumonisin B 1 (FB 1). The assay was based upon the competition between unlabeled FB 1 (i.e. from a sample) and a fluorescein-labeled FB 1 reagent (FB 1-FL). The FB 1-FL was prepared by derivatizing FB 1 with fluorescein isothiocyanate and was purified with affinity columns consisting of a monoclonal antibody (MAb) directed against fumonisins (clone P2A5-3-F3) coupled to agarose. The purified FB 1-FL was subjected to capillary zone electrophoresis. Addition of purified MAb to FB 1-FL before separation resulted in the formation of a complex (MAb.FB 1-FL) with resulting quenching of fluorescence and decrease in the intensity of the FB 1-FL peak. When unlabeled FB 1 was also added to the reaction mixture the FB 1 and FB 1-FL competed for the limited amount of antibody present causing the FB 1-FL peak to increase in direct proportion to the amount of unlabeled FB 1 present. Fumonisin standards could be analyzed with this technique with a total analysis time of 6 min, 2 min of which was required for washing the capillary between analyses. The concentration of unlabeled FB 1 required to obtain 50% of the maximum fluorescence (IC 50) was highly dependent upon the antibody concentration and ranged from 58 to 4170 ng ml-1 at 15-75 µg m-1 of antibody. The optimum performance was seen with 25-50 µg ml-1 of antibody, with IC 50 's between 500 and 1700 ng ml-1 of FB 1. The technique was applied to a limited number of corn samples spiked with high levels of FB 1 (> 10 ppm). This technology holds considerable promise for the rapid analysis of mycotoxins in foods.

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Detoxification of Mycotoxins In Planta as a Strategy for Improving Grain Quality and Disease Resistance: Identification of Fumonisin-Degrading Microbes from Maize

Duvick

Rood

Maddox

et al. 1998

Developments in Plant Pathology

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Immunochemical Methods for Fumonisins

Cited by 6 publications

References 51 publications

Production and Characterization of Monoclonal Antibodies against Fumonisin B1

Production and Characterization of Monoclonal Antibodies against Fumonisin B1

Detection of the mycotoxin fumonisin B₁by a combination of immunofluorescence and capillary electrophoresis

Detoxification of Mycotoxins In Planta as a Strategy for Improving Grain Quality and Disease Resistance: Identification of Fumonisin-Degrading Microbes from Maize

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Immunochemical Methods for Fumonisins

Cited by 6 publications

References 51 publications

Production and Characterization of Monoclonal Antibodies against Fumonisin B1

Production and Characterization of Monoclonal Antibodies against Fumonisin B1

Detection of the mycotoxin fumonisin B1by a combination of immunofluorescence and capillary electrophoresis

Detoxification of Mycotoxins In Planta as a Strategy for Improving Grain Quality and Disease Resistance: Identification of Fumonisin-Degrading Microbes from Maize

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Detection of the mycotoxin fumonisin B₁by a combination of immunofluorescence and capillary electrophoresis