Wang Hai-Qiao, Zhang Hai-Li, Li Xiu-Qing, Wang Jian-Hao, Huang Zhen-Li, Zhao Yuan-Di
Key Laboratory of Biomedical Photonics of Ministry of Education-Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, HuBei 430074, People's Republic of China.
J Biomed Mater Res A. 2008 Sep;86(3):833-41. doi: 10.1002/jbm.a.31682.
Quantum dot (QD) solubilization, conjugation with biomolecules, column purification, and labeling of human HepG2 cells with transferrin-QD (Tf-QD) conjugates are reported in detail in this article. Water-soluble QDs (WQDs) were obtained using sodium thiolycolate to replace the surface ligand tri-n-octylphosphine oxide (TOPO) on the surface of oil-soluble QDs, and Tf-QD conjugates were produced by coupling Tf to WQD. The resulting Tf-QDs were characterized by UV and luminescence spectrophotometry and purified by Sephadex column. The results indicate that Tf has been conjugated to QD successfully. Based on transferrin/transferrin-receptor-mediated delivery system, the Tf-QD conjugates were used to label human HepG2 cells. After a short incubation, the QDs were mainly localized to the membrane of cells. After 12-h incubation, QDs appear mainly in the cytoplasm portion. However, QDs were not found in the nucleus of the cells. Furthermore, the fluorescence intensity of QDs in the cells reduces gradually over time, and fluorescence cannot be observed after 10 days. However, the growth of the labeled cells was not markedly affected by the toxicity of QDs, and they are alive for 10 days. These results can be used for further application of QDs in bioscience.
本文详细报道了量子点(QD)的增溶、与生物分子的共轭、柱纯化以及用转铁蛋白-量子点(Tf-QD)共轭物标记人肝癌细胞系HepG2细胞的过程。通过用硫代乙醇酸钠取代油溶性量子点表面的表面配体三正辛基氧化膦(TOPO)获得水溶性量子点(WQDs),并通过将转铁蛋白与WQDs偶联制备Tf-QD共轭物。所得的Tf-QDs通过紫外和发光分光光度法进行表征,并通过葡聚糖凝胶柱进行纯化。结果表明转铁蛋白已成功与量子点共轭。基于转铁蛋白/转铁蛋白受体介导的递送系统,Tf-QD共轭物用于标记人肝癌细胞系HepG2细胞。短暂孵育后,量子点主要定位于细胞膜。孵育12小时后,量子点主要出现在细胞质部分。然而,在细胞核中未发现量子点。此外,细胞中量子点的荧光强度随时间逐渐降低,10天后无法观察到荧光。然而,标记细胞的生长未受到量子点毒性的明显影响,并且它们存活了10天。这些结果可用于量子点在生物科学中的进一步应用。