Cha Sang-Ho, Kim Sung-Hee, Bischoff Karyn, Kim Hyun-Jeong, Son Seong-Wan, Kang Hwan-Goo
Toxicology and Chemistry Division, Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang 430-824, Korea.
J Vet Sci. 2012 Jun;13(2):119-25. doi: 10.4142/jvs.2012.13.2.119.
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.
使用玉米赤霉烯酮-羧甲基氧胺和-牛血清白蛋白偶联物制备了一种抗玉米赤霉烯酮（ZEN）的单克隆抗体（mAb）。选择了一个显示出非常高结合能力的克隆产生的抗体，发现其与市售的ZEN抗体相比，对ZEN具有更高的亲和力。我们使用所选抗体开发了两种直接竞争ELISA系统（ZEN包被和抗ZEN抗体包被的ELISA）。抗ZEN抗体包被的ELISA和ZEN包被的ELISA的定量范围分别为25至750 ppb和12.5至100 ppb。用标准溶液测定的两种方法的检测限均为10 ppb。两种ELISA的板内和孔间变异均小于10%。与ZEN相比，α-玉米赤霉醇、β-玉米赤霉醇、α-玉米赤霉烯醇和β-玉米赤霉烯醇在ZEN包被的ELISA中的IC(50)值分别为108.1、119.3、114.1和130.3%。在抗ZEN抗体包被的ELISA中，这些值分别为100.7、120.7、121.6和151.6%。根据抗ZEN抗体包被的ELISA，在含有150至600 ng/mL ZEN的加标动物饲料中，ZEN的平均回收率为106.07至123.00%，变异系数为0.93至2.28%。我们的结果表明，本研究中开发的单克隆抗体可用于同时筛选饲料中的ZEN及其代谢物。
