Title: Development and evaluation of monoclonal antibodies for the glucoside of T-2 toxin (T2-Glc) Authors
Submitted to: Toxins
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: July 11, 2013
Publication Date: July 19, 2013
Citation: Maragos, C.M., Kurtzman, C.P., Busman, M., Price, N.P., McCormick, S.P. 2013. Development and evaluation of monoclonal antibodies for the glucoside of T-2 toxin (T2-Glc). Toxins. 5(7):1299-1313. Interpretive Summary: The interactions between fungi and plants can yield metabolites that are toxic to animals. Certain fungi are known to produce a potent toxin known as T-2 toxin. Plants, animals, and fungi protect themselves from toxins through a variety of mechanisms, including glucosylation, which attaches a sugar to the toxin. In general such products are less hazardous than the parent toxins. Unfortunately they can also serve as a ‘reservoir,’ from which the toxin may be regenerated. The glucosylated forms of T-2 toxin have been found in grain and are of interest as potential reservoirs of T-2 toxin that are not detected by many analytical methods. In this research, scientists at the ARS National Center for Agricultural Utilization Research (NCAUR) in Peoria, IL, generated antibodies directed against the glucoside of T-2 toxin. From 10 such antibodies, one was selected that was able to be used in sensitive assays to detect both T-2 toxin and its glucoside. The antibody provides researchers with a new tool that will be used to develop better ways to detect this heretofore ‘masked’ mycotoxin.
Technical Abstract: The interactions between fungi and plants can yield metabolites that are toxic in animal systems. Certain fungi are known to produce sesquiterpenoid trichothecenes, such as T-2 toxin, that are biotransformed by several mechanisms including glucosylation. The glucosylated forms have been found in grain and are of interest as potential reservoirs of T-2 toxin that are not detected by many analytical methods. Hence the glucosides of trichothecenes are often termed “masked” mycotoxins. The glucoside of T-2 toxin (T2-Glc) was linked to keyhole limpet hemocyanin and used to produce antibodies in mice. Ten monoclonal antibody (Mab)-producing hybridoma cell lines were developed. The Mabs were used in immunoassays to detect T2-Glc and T-2 toxin, with midpoints of inhibition curves (IC50s) in the low ng/mL range. Most of the Mabs demonstrated good cross-reactivity to T-2 toxin, with lower recognition of HT-2 toxin. One of the clones (2-13) was further characterized with in-depth cross-reactivity and solvent tolerance studies. Results suggest Mab 2-13 will be useful for the simultaneous detection of T-2 toxin and T2-Glc.