A novel label-free bioengineered cell-based biosensor for salicin detection
A novel label-free bioengineered cell-based biosensor for salicin detection
复制标题
一种用于水杨苷检测的新型无标记生物工程细胞生物传感器
DOI：
10.1016/j.snb.2016.02.072
复制
发表时间：
2017-02-01
影响因子：
8.4
通讯作者：
Wang, Ping
中科院分区：
文献类型：
--
作者：
Hu, Liang;Zou, Lin;Wang, Ping
水杨苷是一种具有重要药用价值的苦味化合物。在制药行业中水杨苷的检测通常采用标准的分析化学方法如高效液相色谱法，由于操作过程复杂，系统昂贵，不适合于一次检测。在这项研究中，我们探讨了使用无标记的生物工程细胞生物传感器检测水杨苷的可行性。以人苦味受体T2 R16为传感元件，在HEK-293细胞中表达水杨苷特异性苦味受体T2 R16。通过免疫荧光染色和钙离子成像验证T2 R16的结构和功能表达。利用细胞阻抗传感器作为二次传感器检测细胞对水杨苷的反应。通过分析不同浓度下水杨苷的响应强度，研究了生物工程细胞对水杨苷的细胞阻抗响应。细胞阻抗测试结果表明，该生物工程细胞对水杨苷的反应具有剂量依赖性。最后，这种生物工程细胞为基础的生物传感器的性能进行了评价。在细胞阻抗测量中，未转染的HEK-293细胞对水杨苷没有响应，这证明了生物传感器的特异性。在标准响应曲线上对两个未测浓度水杨苷的检测也证明了该传感器对水杨苷的检测灵敏度。该方法具有免标记检测的优点，为水杨苷的初步快速检测提供了一种有前途的方法。(C)2016由Elsevier B. V.出版
Salicin is a bitter compound with important pharmaceutical values. The detection of salicin in pharmaceutical industry usually employs standard methods of analytical chemistry such as high performance liquid chromatography, which is not suitable for primary detection due to the complicated operation process and expensive systems. In this study, we explore the feasibility of using a label-free bioengineered cell-based biosensor to detect salicin. A human bitter taste receptor, T2R16, which could specifically respond to salicin, was expressed in HEK-293 cells as sensing elements of the biosensor. The structural and functional expression of T2R16 was validated by immunofluorescent staining and calcium imaging. The cell-impedance sensor was utilized as secondary transducer to detected cellular responses to salicin. The cell-impedance response of the bioengineered cells to salicin was investigated by analyzing the response intensity under various concentrations. The cell-impedance results revealed that this bioengineered cells could respond to salicin in a dose-dependent relationship. Finally, the performance of this bioengineered cell-based biosensor was evaluated. In cell-impedance measurement, non-transfected HEK-293 cells were not responsive to salicin, which proved the specificity of the biosensor. The detection of salicin at two unmeasured concentrations in standard response curve also proved this biosensor was sensitive in salicin detection. With the advantage of label-free measurement, this bioengineered may provide a promising and valuable approach for primary and quick detection of salicin. (C) 2016 Published by Elsevier B.V.