Institute of Biopharmaceutical and Healthcare Engineering, Shenzhen International Graduate School, Tsinghua University, Shenzhen 518055, China.
Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
Analyst. 2022 Jul 12;147(14):3350-3359. doi: 10.1039/d2an00669c.
A sensitive detection system based on aptamer-based biosensors for the detection of carcinoembryonic antigen (CEA) by mapping encoding upconversion nanoparticles (UCNPs) was constructed. In this sensor, oligonucleotides with CEA aptamer fragments immobilized on magnetic beads (MBs) were hybridized to complementary DNA modified on UCNPs (cDNA-UCNPs); thus, sandwich-structured probes were formed. In the presence of CEA, due to the stronger interaction between the aptamer and CEA than that of the aptamer and complementary DNA on UCNPs, the cDNA-UCNPs were isolated from the MBs, and the number of isolated UCNPs was directly related to the concentration of CEA. Using an inverted fluorescence microscope, the number of target-dependent UCNPs on a glass slide was counted, enabling the accurate determination of CEA in the solution. The dynamic range for CEA detection in PBS buffer was 0.02-6.0 ng mL (0.1-30 pM) and a limit of detection (LOD) of 65 fM was achieved. We envisage that the system we developed can also have many promising applications in the sensitive detection of other biomarkers for early cancer diagnosis.
基于适配体的生物传感器的灵敏检测系统,用于通过绘制上转换纳米粒子(UCNPs)的编码来检测癌胚抗原(CEA)。在该传感器中,固定在磁珠(MBs)上的具有 CEA 适配体片段的寡核苷酸与修饰在 UCNPs 上的互补 DNA(cDNA-UCNPs)杂交;从而形成三明治结构的探针。在存在 CEA 的情况下,由于适配体与 CEA 之间的相互作用比适配体与 UCNPs 上的互补 DNA 之间的相互作用更强,因此 cDNA-UCNPs 从 MBs 中分离出来,并且分离的 UCNPs 的数量与 CEA 的浓度直接相关。通过倒置荧光显微镜,对载玻片上的靶标依赖型 UCNPs 的数量进行计数,从而可以准确测定溶液中的 CEA。在 PBS 缓冲液中检测 CEA 的动态范围为 0.02-6.0ng mL(0.1-30 pM),检测限(LOD)达到 65fM。我们预计,我们开发的系统还可以在其他生物标志物的灵敏检测中具有许多有前途的应用,用于早期癌症诊断。