Author
MARAGOS, CHRIS | |
BUSMAN, MARK | |
KONISHI, YOSHIKO - NIH SCIENCES JAPAN |
Submitted to: Journal of Food Additives & Contaminants
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 3/18/2006 Publication Date: 5/1/2006 Citation: Maragos, C.M., Busman, M., Sugita-Konishi, Y. 2006. Production and characterization of a monoclonal antibody cross-reactive with the mycotoxins nivalenol and 4-deoxynivalenol. Food Additives and Contaminants. 23(8):816-825. Interpretive Summary: Nivalenol (NIV) is a mycotoxin produced by certain fungi that are pathogenic to important cereal crops, in particular maize, wheat, and barley. NIV is found worldwide and is closely related to 4-deoxynivalenol (DON or vomitoxin) a mycotoxin associated with outbreaks of Fusarium Head Blight in North America. Literature on the toxicity of NIV suggests it is similar, if not more toxic, than DON. Rapid antibody-based assays for detecting NIV have heretofore not existed without chemically modifying the toxin before analysis. This report describes the development of a monoclonal antibody that can be used to detect both DON and NIV simultaneously at relevant concentrations, without the need to chemically modify either toxin. This antibody will likely find use in rapid assays where simultaneous isolation or detection of both toxins is desirable. Technical Abstract: Nivalenol (NIV) is a mycotoxin produced by certain fungi that are pathogenic to important cereal crops, in particular maize, wheat, and barley. This toxin, 3 alpha,4 beta,7 alpha,15-tetrahydroxy-12,13-epoxytrichothec-9-en-8-one, is found worldwide and is closely related to 4-deoxynivalenol (DON or vomitoxin) a mycotoxin associated with outbreaks of Fusarium Head Blight in North America. Literature on the toxicity of NIV suggests it is similar, if not more toxic, than DON. Despite the development of rapid immunologically-based assays for detecting DON, such assays have not existed for detecting NIV without chemical modification of the analyte. This report describes the development of a monoclonal antibody (Mab) using a NIV-glycine protein conjugate. The Mab is specific for an acetylated form of DON (3-Ac-DON) and cross reacts with both DON and NIV at relevant concentrations without the need to chemically modify the toxin. In a competitive indirect (CI) ELISA format the concentrations of toxins able to inhibit color development by 50% (IC50) were 1.7 ng ml-1, 15.8 ng ml-1, 27.5 ng ml-l, 68.9 ng ml-1 and 1740 ng ml-1 for the mycotoxins 3-Ac-DON, DON, NIV, 15-Ac-DON, and fusarenon-X respectively. The antibody was also used to develop a competitive direct (CD) ELISA for DON and NIV, with IC50's of 16.5 ng ml-1 (DON) and 33.4 ng ml-1 (NIV). These assays are capable of detecting both DON and NIV simultaneously, a property that may be useful in regions where these toxins co-occur or in formats, such as immunoaffinity columns, where co-isolation of both toxins is desirable. |