Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay.
10.4142/jvs.2013.14.2.143
- Author:
Hyuk Mi LEE
1
;
Sung Ok SONG
;
Sang Ho CHA
;
Sung Bok WEE
;
Karyn BISCHOFF
;
Sung Won PARK
;
Seong Wan SON
;
Hwan Goo KANG
;
Myung Haing CHO
Author Information
1. Animal and Plant Quarantine Agency, Anyang 430-757, Korea. kanghg67@korea.kr
- Publication Type:Original Article ; Evaluation Studies ; Research Support, Non-U.S. Gov't
- Keywords:
deoxynivalenol;
ELISA;
magnetic nanoparticles;
monoclonal antibody
- MeSH:
Animal Feed/analysis;
Animals;
Antibodies, Fungal/analysis;
Antibodies, Monoclonal/analysis;
Chemistry Techniques, Analytical/*methods;
Enzyme-Linked Immunosorbent Assay/*methods/veterinary;
Female;
Food Contamination/*analysis;
Fusarium/immunology;
Imidazoles/chemistry;
Magnetics/methods;
Mice;
Mice, Inbred BALB C;
Mycotoxins/*analysis/chemistry;
Nanoparticles/chemistry;
Ovalbumin/chemistry;
Trichothecenes/*analysis/chemistry
- From:Journal of Veterinary Science
2013;14(2):143-150
- CountryRepublic of Korea
- Language:English
-
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 microg/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 microg 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.
