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ARS Home » Midwest Area » Peoria, Illinois » National Center for Agricultural Utilization Research » Mycotoxin Prevention and Applied Microbiology Research » Research » Publications at this Location » Publication #319943
Title: Development and evaluation of monoclonal antibodies for paxilline

Author
item Maragos, Chris

Submitted to: Toxins
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/21/2015
Publication Date: 9/25/2015
Publication URL: https://handle.nal.usda.gov/10113/62296
Citation: Maragos, C.M. 2015. Development and evaluation of monoclonal antibodies for paxilline. Toxins. 7(10):3903-3915.

Interpretive Summary: Paxilline (PAX) is a toxin produced by certain fungi that grow on perennial ryegrass. It is a neurotoxin that causes tremors in animals and, along with a related toxin (Lolitrem B), has been associated with a disease in ruminants known as “ryegrass staggers.” In order to facilitate rapid screening for this toxin, four monoclonal antibodies that bind PAX were developed by a scientist at the ARS National Center for Agricultural Utilization Research (NCAUR) in Peoria, IL. One of the antibodies was used to develop a sensitive method for detecting PAX in maize silage. The assay was applied to 86 silage samples, with only very low levels of PAX detected. Results suggest the method can be applied as a sensitive technique for the rapid screening of PAX in silage. The antibody provides researchers with a new tool that will be used to develop better ways to detect this mycotoxin in plants.

Technical 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 2-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.