Laboratory of Biomedical Engineering, Chongqing Medical University, Chongqing, 400016, People's Republic of China.
Department of Anesthesiology, Affiliated Hospital 1st (Southwest Hospital), Army Medical University, Chongqing, 400038, People's Republic of China.
Mikrochim Acta. 2019 Jun 13;186(7):443. doi: 10.1007/s00604-019-3558-7.
The work describes a simplified method for the preparation of liquid crystal (LC) bioassay using DNA-based capture molecules and having lower detection limits. The capture DNA probes of the stem-loop structure were immobilized on the surface of a glass slide. A homeotropic orientation of LC molecules can be obtained with the proper surface coverage of capture DNA probes. In the presence of analytes (specifically shown here for the progesterone as a model analyte), the molecular binding between capture DNA probes and progesterone opens the loop of the capture DNA probes. The opened sequence is then amenable to hybridization with a reporter DNA probe that is immobilized on gold nanoparticles. This changes the surface microstructure, disrupts the orientation of LC molecules, and results in an enhanced optical response, expressed as the average grey value of the images. This new kind of surface treatment for simultaneous recognition of target molecules and homeotropic anchoring of LCs reduces the number of preparation steps and makes the process of LC bioassay easier. This method has a detection limit as low as 0.1 pmol·L of progesterone. Graphical abstract Schematic presentation of the liquid crystal-based DNA assay. DMOAP: Dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride; TEA: Triethoxsilylbutyraldehyde; 5CB: 4-cyano-4'-pentylbiphenyl; P4: progesterone.
该工作描述了一种使用基于 DNA 的捕获分子制备液晶(LC)生物测定的简化方法,具有更低的检测限。茎环结构的捕获 DNA 探针被固定在玻片表面上。通过适当的捕获 DNA 探针表面覆盖率,可以获得 LC 分子的垂直取向。在存在分析物(这里特别显示为孕激素作为模型分析物)的情况下,捕获 DNA 探针和孕激素之间的分子结合打开了捕获 DNA 探针的环。然后,打开的序列可以与固定在金纳米粒子上的报告 DNA 探针杂交。这会改变表面微观结构,破坏 LC 分子的取向,并导致增强的光学响应,表现为图像的平均灰度值。这种用于同时识别靶分子和 LC 垂直锚定的新型表面处理方法减少了制备步骤的数量,使 LC 生物测定过程更加容易。该方法对孕激素的检测限低至 0.1 pmol·L。