Production of a highly group-specific monoclonal antibody against zearalenone and its application in an enzyme-linked immunosorbent assay.
A monoclonal antibody (mAb) against zearalenone (ZEN) was produced using ZEN-carboxymethoxylamine and -BSA conjugates. Antibody produced by one clone showing a very high binding ability was selected and found to have a higher affinity for ZEN compared to a commerciall ZEN antibody. We developed two direct competitive ELISA systems using the selected antibody (ZEN-coated and anti-ZEN antibody-coated ELISA). Quantitative ranges for the anti-ZEN antibody coated ELISA and ZEN-coated ELISA were from 25 to 750 ppb and from 12.5 to 100 ppb, respectively. The detection limit of both methods as measured with standard solutions was 10 ppb. The intra-plate and inter-well variation of both ELISAs were less than 10%. The IC(50) values for α-zearalenol, β-zearalenol, α-zearalanol, and β-zearalanol compared to ZEN were 108.1, 119.3, 114.1, and 130.3% for the ZEN-coated ELISA. These values were 100.7, 120.7, 121.6, and 151.6% for the anti-ZEN antibody-coated ELISA. According to the anti-ZEN antibody-coated ELISA, the average recovery rates of ZEN from spiked animal feed containing 150 to 600 ng/mL of ZEN ranged from 106.07 to 123.00% with 0.93 to 2.28% coefficients of variation. Our results demonstrate that the mAb developed in this study could be used to simultaneously screen for ZEN and its metabolites in feed.
Toxicology and Chemistry Division, Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang 430-824, Korea.
Sang-Ho
SH
Cha
Sung-Hee
SH
Kim
Karyn
K
Bischoff
Hyun-Jeong
HJ
Kim
Seong-Wan
SW
Son
Hwan-Goo
HG
Kang
D000625:Aminooxyacetic Acid / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D020128:Inhibitory Concentration 50; D051379:Mice; D008807:Mice, Inbred BALB C; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000737:chemistry; D015025:Zearalenone / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000625:Aminooxyacetic Acid; D012710:Serum Albumin, Bovine; D015025:Zearalenone