Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay |
| |
Authors: | Hyuk-Mi Lee Sung-Ok Song Sang-Ho Cha Sung-Bok Wee Karyn Bischoff Sung-Won Park Seong-Wan Son Hwan-Goo Kang Myung-Haing Cho |
| |
Affiliation: | 1.Animal and Plant Quarantine Agency, Anyang 430-757, Korea.;2.Nanobrick Co., Ltd, Suwon 443-270, Korea.;3.Analytical Toxicology Lab, College of Veterinary Medicine, Cornell University, NY 14850, USA.;4.Laboratory of Toxicology, College of Veterinary Medicine, Seoul National University, Seoul 157-742, Korea. |
| |
Abstract: | Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 µg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 µg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs. |
| |
Keywords: | deoxynivalenol ELISA magnetic nanoparticles monoclonal antibody |
|
|