Matos Tiago, Bülow Leif
Pure and Applied Biochemistry, Chemical Center, Lund University, Lund, Sweden.
Curr Protein Pept Sci. 2019;20(1):49-55. doi: 10.2174/1389203718666171024112556.
The needs for purified nucleic acids for preparative and analytical applications have increased constantly, demanding for the development of new and more efficient methods for their recovery and isolation. DNA molecules harbour some intrinsic chemical properties that render them suitable for chromatographic separations. These include a negatively charged phosphate backbone as well as a hydrophobic character originating mainly from the major groove of DNA which exposes the base pairs on the surface of the molecule. In addition, single stranded DNA often allows for a free exposure of the hydrophobic aromatic bases. In this review, multimodal chromatography (MMC) has been evaluated as an alternative tool for complex separations of nucleic acids. MMC embraces more than one kind of interaction between the chromatographic ligand and the target molecules. These resins have often proved superior to conventional single-mode chromatographic materials for DNA isolation, including, e.g., the purification of plasmid DNA from crude cell lysates and for the preparation of DNA fragments before or after a polymerase chain reaction (PCR).
制备和分析应用对纯化核酸的需求不断增加，这就要求开发新的、更高效的核酸回收和分离方法。DNA分子具有一些内在化学性质，使其适合色谱分离。这些性质包括带负电荷的磷酸骨架以及主要源自DNA大沟的疏水性，大沟使碱基对暴露于分子表面。此外，单链DNA常常使疏水性芳香碱基能够自由暴露。在本综述中，多模式色谱法（MMC）已被评估为用于核酸复杂分离的一种替代工具。MMC包含色谱配体与目标分子之间不止一种相互作用。这些树脂在DNA分离方面常常已证明优于传统的单模式色谱材料，包括从粗细胞裂解物中纯化质粒DNA以及在聚合酶链反应（PCR）之前或之后制备DNA片段等。
