Hafez Abdellatif Ahmed Abdelfattah, Abdelhafez Wael Abdellah, Sarhan Hatem Abdelmunsef
Department of Pharmaceutics and Industrial pharmacy, Faculty of Pharmacy, Al-Azhar University, Assuit, Egypt.
Department of Pharmaceutics and Industrial pharmacy, Faculty of Pharmacy, El-Minia University, El-Minia, Egypt.
Iran J Pharm Res. 2018 Spring;17(2):513-524.
Due to the unique optical properties like high brightness and narrow emission bands of Quantum dots, it is used as simple fluorescence materials in bio-imaging, immunoassays, microarrays, and other applications. To easy invistigate cell lines that overexpressed somtostatin receptors, somatostatin (SST) was conjugated with Quantum dots carrying PEG amine (Qdots-PEG-NH). The conjugation of SST to Qdots-PEG-NH started with the thiolation of SST using Traut's reagent. Moreover, the Qdots-PEG-NH were subsequently activated by 500-fold molar excess of sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo-SMCC) dissolved in phosphate buffer. The Qdots-PEG-NH-sulfo-SMCC was conjugated to the thiolated-SST to form Qdots-SST. The number of sulfhydryl groups can be controlled by the molar ratio of Traut´s reagent to SST. Thiolation was necessary for the conjugation of SST to Qdots-PEG-NH. This was achieved by reacting the SST with Traut's reagent in a 1:1 molar ratio. Ellman's reagent was used to determine the number of sulfhydryle groups. Furthermore, cellular uptake study on triple negative breast cancer cells (HCC-1806) showed that the numbers of Qdots-SST per cell were significantly higher compared to unmodified Qdots-PEG-NH when quantified using inductively coupled plasma optical emission spectroscopy (ICP-OES). Moreover, the binding of Qdots-SST to cells can be suppressed by addition of free SST, indicating that the binding of Qdots-SST to cells is due to receptor-specific binding.
由于量子点具有高亮度和窄发射带等独特的光学特性,它被用作生物成像、免疫测定、微阵列及其他应用中的简单荧光材料。为了便于研究过表达生长抑素受体的细胞系,将生长抑素(SST)与携带聚乙二醇胺的量子点(Qdots-PEG-NH)偶联。SST与Qdots-PEG-NH的偶联始于使用特劳特试剂对SST进行硫醇化。此外,随后用溶解在磷酸盐缓冲液中的500倍摩尔过量的磺基琥珀酰亚胺4-(N-马来酰亚胺甲基)环己烷-1-羧酸盐(磺基-SMCC)激活Qdots-PEG-NH。将Qdots-PEG-NH-磺基-SMCC与硫醇化的SST偶联以形成Qdots-SST。巯基的数量可以通过特劳特试剂与SST的摩尔比来控制。硫醇化对于SST与Qdots-PEG-NH的偶联是必要的。这通过使SST与特劳特试剂以1:1的摩尔比反应来实现。使用埃尔曼试剂来确定巯基的数量。此外,对三阴性乳腺癌细胞(HCC-1806)的细胞摄取研究表明,当使用电感耦合等离子体发射光谱法(ICP-OES)进行定量时,每个细胞中Qdots-SST的数量比未修饰的Qdots-PEG-NH显著更高。此外,添加游离SST可以抑制Qdots-SST与细胞的结合,表明Qdots-SST与细胞的结合是由于受体特异性结合。