a Iran National Science Foundation: INSF , Iran.
b Department of Medical Biotechnology , Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences , Tabriz , Iran.
Artif Cells Nanomed Biotechnol. 2016 Sep;44(6):1417-23. doi: 10.3109/21691401.2015.1031905. Epub 2015 Apr 17.
Aimml: The aim of this project was to synthesize and characterize gold nanoparticles (GNPs) to trace the sequence of the hnRNPB1as a lung cancer biomarker.
In the synthesis of GNPs with characteristics appropriate for conjugation, the size, morphology, and shape of the synthesized GNPs were determined by using spectrophotometry and transmission electron microscopy (TEM), followed by designing a probe for hnRNPB1biomarker with characteristics suitable for conjugation. Next, the GNPs were functionalized with a single-stranded DNA probe that was specific for the biomarker, for the characterization and confirmation of the conjugation process. Finally, for determination of minimum level of detection in solution including DNA target and probe aggregation, the changes in the absorption spectra of the samples in the range of 250-750 nm were determined using the NanoDrop ND 1000 spectrophotometer.
The surface of GNPs can be modified by utilizing ligands to selectively attach biomarkers. Thiol-bonding of DNA and chemical functionalization of GNPs are the most common approaches. Colloidal gold was synthesized with the citrate reduction method, as described by Turkevich et al. in 1951. In this study, the probe for hnRNPB1 was designed with a thiol crosslinker. Every set of conjugated GNPs was complementary to one end of the hnRNPB1 biomarker, and the probes were aligned in a tail to tail fashion onto the target.
Uniform GNPs were synthesized by the citrate reduction technique, and the outcomes of trials with variation in factors (shape and size of the nanoparticles, gold concentration, and conjugation between GNP and probes) were investigated. The gold nanoprobe-based technique is better than the PCR-based techniques, because there are no requirements of enzymatic amplification and gel electrophoresis, and the evaluation can be done using small amounts of sample.
本项目旨在合成和表征金纳米粒子（GNPs），以追踪 hnRNPB1 作为肺癌生物标志物的序列。
在合成具有适当共轭特征的 GNPs 时，通过分光光度法和透射电子显微镜（TEM）确定合成 GNPs 的大小、形态和形状，然后设计适用于共轭的 hnRNPB1 生物标志物的探针。接下来，将 GNPs 功能化，使其与特定于生物标志物的单链 DNA 探针结合，用于共轭过程的表征和确认。最后，为了确定溶液中包括 DNA 靶标和探针聚集在内的最小检测限，使用 NanoDrop ND 1000 分光光度计测定样品在 250-750nm 范围内的吸收光谱变化。
可以通过利用配体来修饰 GNPs 的表面，以选择性地连接生物标志物。DNA 的硫醇键合和 GNPs 的化学功能化是最常见的方法。胶体金是通过 Turkevich 等人在 1951 年描述的柠檬酸还原法合成的。在本研究中，设计了用于 hnRNPB1 的探针带有硫醇交联剂。每一组共轭 GNPs 都与 hnRNPB1 生物标志物的一端互补，并且探针以尾对尾的方式排列在靶标上。
通过柠檬酸还原技术合成了均匀的 GNPs，并研究了不同因素（纳米颗粒的形状和大小、金浓度以及 GNP 和探针之间的共轭）的试验结果。基于金纳米探针的技术优于基于 PCR 的技术，因为不需要酶扩增和凝胶电泳，并且可以使用少量样品进行评估。
