| 应用基于单克隆抗体的免疫传感器检测环境中的芘和苯并芘 |
| 摘要点击 1512 全文点击 907 投稿时间：2015-03-04 修订日期：2015-06-29 |
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| 中文关键词 苯并芘 芘 单克隆抗体 免疫检测 传感器 |
| 英文关键词 pyrene benzo(a)pyrene monoclonal antibody immunoassay sensor |
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| 中文摘要 |
| 多环芳烃是一类普遍存在的环境污染物,可以诱发癌症,对人们的健康造成严重威胁. 本研究利用一种四环芳烃芘的衍生物作为半抗原,利用活泼酯法合成人工抗原免疫小鼠研制多环芳烃单克隆抗体. 通过非竞争性ELISA与竞争性ELISA相结合的筛选方式得到一株单克隆抗体,命名为6A6. 该抗体能特异性识别芘和苯并芘,其IC50值分别为8.1 μg ·L-1和6.8 μg ·L-1,对其他多环芳烃的交叉反应率均低于5%. 在此基础上,建立了免疫传感器法对芘和苯并芘进行检测,线性范围为0.2~10.0 μg ·L-1. 利用该方法对添加在水样中的样品进行回收,并与传统ELISA方法进行比对,结果证明传感器法检测更加快速且灵敏度更高. 该方法可以用于含有芘和苯并芘的环境样品的快速筛选,可以在10 min内初步获得检测结果. |
| 英文摘要 |
| Polycyclic aromatic hydrocarbons (PAHs) are common environmental pollutants which are concerned because several of its members have been known to be carcinogenic and mutagenic. Pyrene, as a kind of PAHs, was covalently linked to carrier protein bovine serum albumin by active ester method and immunized to mice. A sensitive mAb 6A6 was developed by a three-step screening procedure. The IC50 of 6A6 against pyrene and benzo(a)pyrene were 8.1 μg ·L-1and 6.8 μg ·L-1, respectively. The cross-reactivity values (CR) with other PAHs were less than 5%. An immunosensor was developed based on 6A6 for pyrene and benzo(a)pyrene detection with a linear range of 0.2-10.0 μg ·L-1. The assessment of spiked water samples by the sensor demonstrated faster and more sensitive than ELISA method. We believe that this sensor can be used to rapidly assess pyrene and benzo(a)pyrene in environmental samples. |
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