Klepárník Karel, Datinská Vladimíra, Voráčová Ivona, Lišková Marcela
Institute of Analytical Chemistry, Czech Academy of Sciences, Veveří 97, 602 00, Brno, Czech Republic,
Methods Mol Biol. 2014;1199:33-54. doi: 10.1007/978-1-4939-1280-3_3.
In many bioanalytical applications, important molecules such as DNA, proteins, and antibodies are routinely conjugated with fluorescent tags to reach an extraordinary sensitivity of analyses. Semiconductor nanoparticles, quantum dots, have already proved to be suitable components of highly luminescent tags, probes, and sensors with a broad applicability in analytical chemistry. Quantum dots provide high extinction coefficients together with a wide range of excitation wavelengths, size- and composition-tunable emissions, narrow and symmetric emission spectra, good quantum yields, relatively long size-dependent luminescence lifetime, and practically no photobleaching. Most of these properties are superior when compared with conventional organic fluorescent dyes. In this chapter, optimized procedures for the preparation of water-dispersed cadmium telluride (CdTe) quantum dots, conjugating reactions with antibodies, DNA, and macrocycles as well as their analyses by capillary electrophoresis are described. The potential of capillary electrophoresis for fast analyses of nanoparticles, their conjugates with antibodies, and immunocomplexes with targeted antigens is demonstrated on examples.
在许多生物分析应用中,DNA、蛋白质和抗体等重要分子通常会与荧光标签缀合,以实现极高的分析灵敏度。半导体纳米颗粒,即量子点,已被证明是高发光标签、探针和传感器的合适组件,在分析化学中具有广泛的适用性。量子点具有高消光系数、宽激发波长范围、尺寸和组成可调的发射、窄且对称的发射光谱、良好的量子产率、相对较长的与尺寸相关的发光寿命,并且几乎没有光漂白现象。与传统有机荧光染料相比,这些特性大多更为优越。本章描述了制备水分散性碲化镉(CdTe)量子点的优化程序、与抗体、DNA和大环化合物的缀合反应以及通过毛细管电泳进行的分析。通过实例展示了毛细管电泳在快速分析纳米颗粒、其与抗体的缀合物以及与靶向抗原的免疫复合物方面的潜力。