Author/Authors :
Tomoyuki Kadota، نويسنده , , Yoko Takezawa، نويسنده , , Satoshi Hirano، نويسنده , , Osamu Tajima، نويسنده , , Chris M. Maragos، نويسنده , , Takashi Nakajima، نويسنده , , Toshitsugu Tanaka، نويسنده , , Yoichi Kamata c، نويسنده , , Yoshiko Sugita-Konishi، نويسنده ,
Abstract :
A surface plasmon resonance (SPR) immunoassay using a monoclonal antibody was developed to measure nivalenol (NIV) and deoxynivalenol (DON) contamination in wheat. A highly sensitive and stable DON-immobilized sensor chip was prepared, and an SPR detection procedure was developed. The competitive inhibition assay used a monoclonal antibody that cross-reacts with NIV and DON. The half maximal inhibitory concentration (IC50) values of the SPR assay were 28.8 and 14.9 ng mL−1 for NIV and DON, respectively. The combined responses of NIV and DON in wheat were obtained using a simultaneous detection assay in a one-step cleanup procedure. NIV and DON were separated using a commercial DON-specific immunoaffinity column (IAC) and their responses were obtained using an independent detection assay. Spiked tests using these toxins revealed that recoveries were in the range 91.5–107% with good relative standard deviations (RSDs) (0.40–4.1%) and that detection limits were 0.1 and 0.05 mg kg−1 for NIV and DON, respectively. The independent detection using IAC showed detection limits of 0.2 and 0.1 mg kg−1 for NIV and DON, respectively. SPR analysis results were correlated with those obtained using a conventional LC/MS/MS method for wheat co-contaminated with NIV and DON. These results suggested that the developed SPR assay is a practical method to rapidly screen the NIV and DON co-contamination of wheat and one of a very few immunoassays to detect NIV directly.