Chemical Sciences Division, Saha Institute of Nuclear Physics , 1/AF Bidhannagar, Kolkata 700064, India.
Institute of Environmental Engineering, National Chiao Tung University , 1001 University Road, Hsinchu 30010, Taiwan.
Anal Chem. 2017 Nov 21;89(22):12244-12251. doi: 10.1021/acs.analchem.7b03095. Epub 2017 Nov 10.
The interest in DNA-protein-based diagnostics has recently been growing enormously, which makes the separation process of DNA or protein from a cell extract extremely important. Unlike the traditional separation process, a novel approach is in demand which can nondestructively isolate the target biomolecules without sacrificing the other components in the mixture. In this study, we have demonstrated a new and simple separation technique by using well-established bifunctional Au-FeO nanocomposites as the separation nanoprobes to efficiently isolate the specifically targeted nanomolar concentrated DNA over 70% from its associate DNA-protein mixture in the presence of a magnetic field. The sensing accuracy of both as-separated DNA and protein are quantitatively examined by UV-vis spectroscopy, and then qualitatively validated by gel analysis. Results obtained in this study clearly demonstrated that this newly developed separation procedure not only provides the efficient separation for the targeted DNA but can also maintain the bioactivity of as-separated protein and DNA solutions. The superiority of this technique can open an avenue to establish a label-free and nondestructive platform for a wide variety of biomolecule separation applications.
基于 DNA-蛋白质的诊断方法近来受到了极大的关注，这使得从细胞提取物中分离 DNA 或蛋白质的过程变得非常重要。与传统的分离过程不同，人们需要一种新的方法，能够在不牺牲混合物中其他成分的情况下，非破坏性地分离目标生物分子。在这项研究中，我们展示了一种新的简单分离技术，使用成熟的双功能 Au-FeO 纳米复合材料作为分离探针，在磁场的存在下，从其相关的 DNA-蛋白质混合物中高效分离出特异性靶向的纳摩尔浓度 DNA，效率超过 70%。通过紫外-可见光谱定量检测了分离出的 DNA 和蛋白质的传感准确性，并通过凝胶分析进行了定性验证。本研究的结果清楚地表明，这种新开发的分离程序不仅可以为目标 DNA 提供高效的分离，还可以保持分离出的蛋白质和 DNA 溶液的生物活性。该技术的优越性为建立各种生物分子分离应用的无标记和非破坏性平台开辟了道路。
