Ashtari Parviz, He Xiaoxiao, Wang Kemin, Gong Ping
Biomedical Engineering Center, State Key Laboratory of Chemo/Biosensing and Chemometrics, Institute of Life Science and Biological Technology, Hunan University & Engineering Research Center for Bio-Nanotechnology of Hunan Province, Changsha 410082, PR China.
Talanta. 2005 Sep 15;67(3):548-54. doi: 10.1016/j.talanta.2005.06.043. Epub 2005 Aug 1.
In this paper, an improved recovery method for target ssDNA using amino-modified silica-coated magnetic nanoparticles (ASMNPs) is reported. This method takes advantages of the amino-modified silica-coated magnetic nanoparticles prepared using water-in-oil microemulsion technique, which employs amino-modified silica as the shell and iron oxide as the core of the magnetic nanoparticles. The nanoparticles have a silica surface with amino groups and can be conjugated with any desired bio-molecules through many existing amino group chemistry. In this research, a linear DNA probe was immobilized onto nanoparticles through streptavidin conjugation using covalent bonds. A target ssDNA(I) (5'-TMR-CGCATAGGGCCTCGTGATAC-3') has been successfully recovered from a crude sample under a magnet field through their special recognition and hybridization. A designed ssDNA fragment of severe acute respiratory syndrome (SARS) virus at a much lower concentration than the target ssDNA(I) was also recovered with high efficiency and good selectivity.
本文报道了一种使用氨基修饰的二氧化硅包覆磁性纳米颗粒(ASMNPs)回收目标单链DNA的改进方法。该方法利用了通过油包水微乳液技术制备的氨基修饰的二氧化硅包覆磁性纳米颗粒,其以氨基修饰的二氧化硅为壳,以氧化铁为磁性纳米颗粒的核。这些纳米颗粒具有带有氨基的二氧化硅表面,并且可以通过许多现有的氨基化学方法与任何所需的生物分子缀合。在本研究中,通过使用共价键的链霉亲和素缀合将线性DNA探针固定在纳米颗粒上。通过它们的特异性识别和杂交,已在磁场下从粗样品中成功回收了目标单链DNA(I)(5'-TMR-CGCATAGGGCCTCGTGATAC-3')。还以高效率和良好的选择性回收了一种设计的严重急性呼吸综合征(SARS)病毒单链DNA片段,其浓度远低于目标单链DNA(I)。