title: Development and Evaluation of Monoclonal Antibodies for Paxilline.
abstract: Paxilline (PAX) is a tremorgenic mycotoxin that has been found in perennial ryegrass infected with Acremonium lolii. To facilitate screening for this toxin, four murine monoclonal antibodies (mAbs) were developed. In competitive indirect enzyme-linked immunosorbent assays (CI-ELISAs) the concentrations of PAX required to inhibit signal development by 50% (IC50s) ranged from 1.2 to 2.5 ng/mL. One mAb (2-9) was applied to the detection of PAX in maize silage. The assay was sensitive to the effects of solvents, with 5% acetonitrile or 20% methanol causing a two-fold or greater increase in IC50. For analysis of silage samples, extracts were cleaned up by adsorbing potential matrix interferences onto a solid phase extraction column. The non-retained extract was then diluted with buffer to reduce solvent content prior to assay. Using this method, the limit of detection for PAX in dried silage was 15 µg/kg and the limit of quantification was 90 µg/kg. Recovery from samples spiked over the range of 100 to 1000 µg/kg averaged 106% ± 18%. The assay was applied to 86 maize silage samples, with many having detectable, but none having quantifiable, levels of PAX. The results suggest the CI-ELISA can be applied as a sensitive technique for the screening of PAX in maize silage.
authors.affiliation: Bacterial Foodborne Pathogens and Mycology Research Unit, USDA-ARS-NCAUR, 1815 North University Street, Peoria, IL 61604, USA. chris.maragos@ars.usda.gov.
authors.forename: Chris M
authors.initials: CM
authors.lastname: Maragos
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived; D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007211:Indoles / Q000032:analysis* / Q000276:immunology; D057230:Limit of Detection; D051379:Mice; D009183:Mycotoxins / Q000032:analysis* / Q000276:immunology; D012015:Reference Standards; D003313:Zea mays / Q000382:microbiology
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D007211:Indoles; D009183:Mycotoxins; C048220:paxilline
keywords: antibody; immunoassay; mycotoxin; paxilline; silage; tremorgen