Analysis of the Interaction between Tropomyosin Allergens and Antibodies Using a Biosensor Based on Imaging Ellipsometry
Analysis of the Interaction between Tropomyosin Allergens and Antibodies Using a Biosensor Based on Imaging Ellipsometry
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DOI：
10.1021/ac303783j
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发表时间：
2013-05-07
影响因子：
7.4
通讯作者：
Chen, Ying
中科院分区：
文献类型：
--
作者：
Wang, Wei;Qi, Cai;Chen, Ying
研究了基于高空间分辨率成像椭圆光度法的生物传感器，以检验其在快速检测和分析甲壳类动物中存在的原肌球蛋白过敏原中的作用。该方法已建立用于检测原肌球蛋白过敏原，包括配体筛选、灵敏度测定以及与传统检测方法的比较。筛选出已知对原肌球蛋白过敏原具有高生物活性的三种单克隆抗体（2F9、4C7和2H6），并将其作为配体分别固定在硅片表面，从而使它们能够捕获原肌球蛋白过敏原。晶片表面产生的变化通过椭圆光度图像上的灰度变化进行可视化。图像显示，这些抗体能够识别虾和蟹中原肌球蛋白过敏原的存在，灵敏度为 1 mg L-1，提取样品的检测时间约为 30 分钟。这项初步研究表明，当对含有虾和蟹的产品使用这种类型的检测时，原肌球蛋白过敏原的检测是快速且特异的。与酶联免疫吸附测定相比，基于成像椭圆光度法的生物传感器能够进行快速的无标记分析。
A biosensor based on high spatial resolution imaging ellipsometry has been studied to examine its role in the rapid detection and analysis of the tropomyosin allergen existing in crustaceans. This methodology has been established for detection of the tropomyosin allergen and includes ligand screening, the determination of sensitivity, and a comparison with traditional detection methods. Three kinds of monoclonal antibodies (2F9, 4C7, and 2H6) known to have a high bioactivity against the tropomyosin allergen were screened and separately immobilized as ligands on a silicon wafer surface, thus allowing them to capture the tropomyosin allergen. Resulting changes on the wafer surface were visualized in gray scale variation on an ellipsometry image. Images showed that these antibodies are able to recognize the presence of the tropomyosin allergen in shrimp and crab with sensitivity of 1 mg L-1 and at a detecting time of approximately 30 min for an extracted sample. This preliminary study has shown that the detection of the tropomyosin allergen is rapid and specific when using this type of assay on products containing shrimp and crab. When compared with the enzyme linked immunosorbent assay, the biosensor based on imaging ellipsometry is able to perform a fast label-free analysis.