DEVELOPMENT AND EVALUATION OF IMPROVED MATERIALS FOR MYCOTOXIN ANALYSIS
Location: Bacterial Foodborne Pathogens & Mycology Research Unit
Title: Rapid Screening Assay for the Detection of Nivalenol and Deoxynivalenol using Monoclonal Antibody and Surface Plasmon Resonance
| Kadota, Tomoyuki - KIRIN HOLDINGS CO LTD |
| Ito, Yuji - KIRIN HOLDINGS CO LTD |
| Takezawa, Yoko - KIRIN HOLDINGS CO LTD |
| Nakajima, Takashi - NATL AG RSCH OKINAWA |
| Tanaka, Toshitsugu - KOBE INST OF HLTH |
| Konishi, Yoshiko-Sugita - NATL INST OF HLTH SCI |
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: November 18, 2008
Publication Date: November 17, 2008
Citation: Kadota, T., Ito, Y., Takezawa, Y., Maragos, C.M., Nakajima, T., Tanaka, T., Konishi, Y. 2008. Rapid Screening Assay for the Detection of Nivalenol and Deoxynivalenol using Monoclonal Antibody and Surface Plasmon Resonance [abstract]. World Mycotoxin Forum. p. 64.
Nivalenol (NIV) and Deoxynivalenol (DON) are trichothecene mycotoxins produced by Fusarium spp that contaminate mainly cereal crops, such as wheat, barley, and maize. These mycotoxins are serious health hazards to human and domestic animals. The study reports a rapid screening method of NIV and DON in wheat using a monoclonal antibody that reacts with both NIV and DON , and a Surface Plasmon Resonance (SPR) biosensor based upon antigen-antibody interaction. SPR assay was performed using a Biacore T100 SPR system (GE Healthcare Bio-Science) with DON-BSA conjugate immobilized on the sensor chip. An Inhibition assay using DON-BSA conjugate on the sensor chip and antibody was developed, and calibration curves had a good correlation over the range of 1.0-25 ng/ml (NIV) and 0.5-25 ng/ml (DON). After each assay, guanidine hydrochloride in glycine solution was injected to the sensor chip to dissociate antibody from DON-BSA. The main steps of sample preparation for wheat were simply extraction, centrifugation, and filtration. Wheat samples were extracted with water for 3 min, and the extract was centrifuged at 10000 rpm for 1 min with a mini centrifuge. The supernatant solution was diluted with HBS-P+ buffer, filtered, and injected to SPR system. The test solution was mixed with antibody using the autosampler of the SPR system. In the case of the SPR assay for NIV, the extract after centrifugation was passed through an immunoaffinity column to remove DON in the test solution. Results of recovery tests for spiked NIV and DON at levels of 100-1000 ng/g and 50-1000 ng/g were 91.9-103% (NIV) and 93.6-118% (DON). In the case of wheat, the detection limit of this assay was 100 ng/g (NIV) and 50 ng/g (DON). Analyses of naturally contaminated wheat using SPR assay were achieved. By using a suitable calibration curve of NIV and DON, the measured values obtained by SPR corresponded to values obtained by LC/MS. In this assay, concentration values analyzed with SPR become the total value of NIV and DON. However, the SPR assay using immunoaffinity column for DON indicated the possibility that NIV can be analyzed by excluding the influence of DON.