复杂样品中玉米赤霉烯酮免疫亲和柱酶免疫分析方法的建立与应用
Development and application of an immunoaffinity column enzyme immunoassay for mycotoxin zearalenone in complicated samples.
作者信息
Tang Xiaoqian, Li Xin, Li Peiwu, Zhang Qi, Li Ran, Zhang Wen, Ding Xiaoxia, Lei Jiawen, Zhang Zhaowei
机构信息
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China ; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China ; Key laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China ; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, P. R. China.
Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan, P. R. China ; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, P. R. China ; Key laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, P. R. China ; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan, P. R. China ; Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan, P. R. China.
出版信息
PLoS One. 2014 Jan 17;9(1):e85606. doi: 10.1371/journal.pone.0085606. eCollection 2014.
The zearalenone (ZEA) monoclonal antibody (mAb) 2D3, one of the highest sensitivity antibodies, was developed. Based on this mAb, it was established of an immunoaffinity column (IAC) coupled with an indirect competitive enzyme-linked immunosorbent assay (icELISA). After optimization, the icELISA allowed an IC50 against ZEA of 0.02 µg L(-1). The mAb 2D3 exhibited a high recognition of ZEA (100%) and β-zearalenol (β-ZOL, 88.2%). Its cross-reactivity with α-zearalenol (α-ZOL) and β-zearalanol (β-ZAL) were found to be 4.4% and 4.6%, respectively. The IAC-icELISA method was employed to analyze ZEA contamination in food samples, compared with high-performance liquid chromatography (HPLC). The spiked assay for ZEA demonstrated the considerable recoveries for IAC-icELISA (83-93%) and HPLC (94-108%) methods. Results showed that the mAb 2D3 and IAC-icELISA method posed potential applications in sensitively determination of ZEA in maize.
开发了最高灵敏度抗体之一的玉米赤霉烯酮（ZEA）单克隆抗体（mAb）2D3。基于该单克隆抗体，建立了一种与间接竞争酶联免疫吸附测定（icELISA）相结合的免疫亲和柱（IAC）。经过优化，icELISA对ZEA的IC50为0.02 µg L(-1)。单克隆抗体2D3对ZEA（100%）和β-玉米赤霉醇（β-ZOL，88.2%）表现出高度识别。发现其与α-玉米赤霉醇（α-ZOL）和β-玉米赤霉烯醇（β-ZAL）的交叉反应率分别为4.4%和4.6%。采用IAC-icELISA方法与高效液相色谱法（HPLC）比较分析食品样品中的ZEA污染。ZEA的加标试验表明，IAC-icELISA方法（83-93%）和HPLC方法（94-108%）具有相当高的回收率。结果表明，单克隆抗体2D3和IAC-icELISA方法在灵敏测定玉米中的ZEA方面具有潜在应用。
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