Huang Xiangyi, Weng Jifang, Sang Fuming, Song Xingtao, Cao Chengxi, Ren Jicun
College of Chemistry & Chemical Engineering, Shanghai Jiaotong University, 800 Dongchuan Road, Shanghai 200240, China.
J Chromatogr A. 2006 Apr 28;1113(1-2):251-4. doi: 10.1016/j.chroma.2006.02.087. Epub 2006 Mar 24.
In this paper, we present a universal, highly efficient and sensitive method for the characterization of quantum dot (QD) bioconjugates based on capillary electrophoresis with laser-induced fluorescent (LIF) detection. We first prepared CdTe QDs in aqueous phase by a chemical route with mercaptopropionic acid as a ligand, and then were coupled to certain proteins using bifunctional linkage reagent or electrostatic attraction. The QD bioconjugates were characterized by capillary electrophoresis with LIF detection. We found that QD bioconjugates were efficiently separated with free QDs by the optimization of buffer pH. Furthermore, we found that ultrafiltration was an effective and simple approach to purify QD conjugates with bovine serum albumin (BSA). Due to their broad absorption spectra and size dependent emission wavelength tunability, QDs can be excited to emit different colour fluorescence using a single wavelength laser source, and therefore, we believe that CE with LIF detection will become a universal and efficient tool for the characterization of QD bioconjugates.
在本文中,我们提出了一种基于毛细管电泳结合激光诱导荧光(LIF)检测的通用、高效且灵敏的量子点(QD)生物共轭物表征方法。我们首先通过化学方法以巯基丙酸作为配体在水相中制备了碲化镉量子点,然后使用双功能连接试剂或静电引力将其与特定蛋白质偶联。通过毛细管电泳结合LIF检测对量子点生物共轭物进行了表征。我们发现,通过优化缓冲液pH值,量子点生物共轭物能与游离量子点有效分离。此外,我们发现超滤是一种有效且简便的方法,可用于纯化与牛血清白蛋白(BSA)结合的量子点共轭物。由于量子点具有宽吸收光谱以及发射波长随尺寸可调谐的特性,使用单一波长激光源即可激发量子点发射不同颜色的荧光,因此,我们认为毛细管电泳结合LIF检测将成为量子点生物共轭物表征的通用且高效的工具。