Production and characterization of monoclonal antibodies against ochratoxin B. |
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| Authors: | Alexandra H Heussner Ines Moeller Billy W Day Daniel R Dietrich Evelyn O'Brien |
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| Affiliation: | Environmental Toxicology, University of Konstanz, P.O. Box X-918, 78457 Konstanz, Germany. |
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| Abstract: | Monoclonal antibodies against ochratoxin B (OTB) were generated by immunizing Balb/c mice with OTB conjugated to keyhole limpet hemocyanin (KLH) via carbodiimide reactions with CHMC and EDAC. A stable hybridoma cell line 2F1.E10 was produced by fusion of murine splenocytes and myeloma cells. The obtained antibodies were characterized using an indirect competitive ELISA. The detection limit was calculated (27+/-2 nM OTB) and 50% binding inhibition was reached at 500 nM free OTB. A low cross-reactivity to ochratoxin A (OTA) of 3.3% and no cross-reactivities to either coumarin or DL-phenylalanine were observed, suggesting a highly specific OTB antibody. The antibody type was identified as IgG class 1 with the light chain being of the kappa configuration. These antibodies can be used in an indirect competitive ELISA to detect OTB in the nanomolar to micromolar concentration range and may be useful for the analysis of contaminated food items. |
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| Keywords: | ELISA, enzyme-linked immunosorbent assay HPLC, high performance liquid chromatography FBS, fetal bovine serum DMSO, dimethylsulfoxide PBS, phosphate-balanced saline PCR, polymerase chain reaction BSA, bovine serum albumin IgG, immunglobulin G IC50, 50% inhibitory concentration RIA, radioimmunoassay |
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