School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, PR China.
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, PR China.
Talanta. 2018 May 15;182:306-313. doi: 10.1016/j.talanta.2018.01.067. Epub 2018 Feb 3.
A novel dual-target recognition sandwich strategy for selective capture and detection of MCF-7 breast cancer cells based on core-shell magnetic mesoporous silica (FeO@nSiO@mSiO@apt) nanoparticles was developed. FeO@nSiO@mSiO@apt nanoparticles, which were prepared by a layer-by-layer method and were used for the first time to capture cancer cells, have large surface areas, particularly accessible mesochannels, and good biocompatibility, enabling aptamers to be compactly anchored onto the surface of the core-shell magnetic nanoparticles. A mucin 1 protein (MUC1)-targeted FeO@nSiO@mSiO@apt nanoparticle was used as an affinity magnetic isolate material to capture target MCF-7 cells selectively and to reduce interference through affinity interaction between the anti-MUC1 aptamer and the MUC1 protein over-expressed on the surface of the MCF-7 cells. Meanwhile, a folate receptor (FR)-targeted affinity fluorescent probe (FA-BSA-FITC) was developed by coupling folic acid and FITC to the surface of BSA, enabling high sensitivity, selective fluorescent labeling of FR over-expressed MCF-7 cells. A dual-target recognition sandwich assay was developed based on the MUC1-targeted magnetic nanoparticles and the FR-targeted fluorescent probes. Under optimum conditions, a quantitative assay of MCF-7 cells was achieved with a dynamic range of 10-10 cells/mL (R = 0.9991). This assay showed high specificity and sensitivity to the target MCF-7 cells. Finally, the proposed strategy could be extended to detect MCF-7 cells in human plasma and whole blood with a recovery range of 86.1-104.0% and a RSD range of 1.2-8.4%, respectively. This indicates that the dual-target recognition method developed in this research exhibits good selectivity, anti-interference capability, and reliability even in plasma and whole blood samples and is more suitable for complex samples than previous targeted assays. Therefore, the approach proposed here may have great potential for early breast cancer diagnosis.
基于核壳型磁性介孔硅（FeO@nSiO@mSiO@apt）纳米粒子，开发了一种新颖的双重靶标识别夹心策略，用于选择性捕获和检测 MCF-7 乳腺癌细胞。FeO@nSiO@mSiO@apt 纳米粒子是通过层层方法制备的，这是它们首次被用于捕获癌细胞。这些纳米粒子具有较大的表面积、特别可及的介孔道和良好的生物相容性，使得适配体能够紧凑地锚定在核壳磁性纳米粒子的表面。一种粘蛋白 1 蛋白（MUC1）靶向的 FeO@nSiO@mSiO@apt 纳米粒子被用作亲和磁分离材料，以选择性地捕获靶标 MCF-7 细胞，并通过抗 MUC1 适配体与 MCF-7 细胞表面过表达的 MUC1 蛋白之间的亲和相互作用来减少干扰。同时，通过将叶酸和 FITC 偶联到 BSA 的表面，开发了一种叶酸受体（FR）靶向的亲和荧光探针（FA-BSA-FITC），使得 FR 过表达的 MCF-7 细胞能够进行高灵敏度、选择性的荧光标记。基于 MUC1 靶向磁性纳米粒子和 FR 靶向荧光探针，建立了一种双重靶标识别夹心测定法。在最佳条件下，实现了 MCF-7 细胞的定量检测，其动态范围为 10-10 细胞/mL（R = 0.9991）。该测定法对靶标 MCF-7 细胞具有高特异性和灵敏度。最后，该策略可扩展用于检测人血浆和全血中的 MCF-7 细胞，回收率范围为 86.1-104.0%，RSD 范围为 1.2-8.4%。这表明，即使在血浆和全血样本中，本研究中开发的双重靶标识别方法也表现出良好的选择性、抗干扰能力和可靠性，并且比以前的靶向测定法更适合复杂样本。因此，这里提出的方法在早期乳腺癌诊断中可能具有很大的潜力。
