Sanandaji Nima, Carlsson Nils, Voinova Marina, Akerman Björn
Department of Chemistry and Bioscience, Chalmers University of Technology, S-41296 Göteborg, Sweden.
Electrophoresis. 2006 Aug;27(15):3007-17. doi: 10.1002/elps.200500812.
Porous hydrogels such as agarose are commonly used to analyze DNA and water-soluble proteins by electrophoresis. More recently lyotropic liquid crystals, such as the diamond cubic phase formed by the lipid monoolein and water, has become a new type of well-defined porous structure of interest for both hydrophilic and amphiphilic analytes. Here we compare these two types of matrixes by investigating the nature of retardation they confer to an oligonucleotide that migrates in their respective aqueous phases. The retardation for a 25-mer oligonucleotide was found to be about 35-fold stronger in the cubic phase than in an agarose hydrogel modified to have the same average pore size. According to modelling, the strong retardation is primarily due to the fact that hydrodynamic interaction with the continuous monoolein membrane is a stronger source of friction than the steric interactions (collisions) with discrete gel fibres. A secondary effect is that the regular liquid crystal has a narrower pore-size distribution than the random network of the agarose gel. In agreement with experiments, these two effects together predict that the retardation in the cubic phase is a 30-fold stronger than in an agarose gel with the same average pore radius.
诸如琼脂糖之类的多孔水凝胶通常用于通过电泳分析DNA和水溶性蛋白质。最近,溶致液晶,例如由单油酸甘油酯和水形成的金刚石立方相,已成为一种新型的具有明确结构的多孔结构,对亲水性和两亲性分析物都很有吸引力。在这里,我们通过研究它们赋予在各自水相中迁移的寡核苷酸的阻滞性质,来比较这两种类型的基质。发现对于一个25聚体寡核苷酸,其在立方相中的阻滞作用比在经修饰以具有相同平均孔径的琼脂糖水凝胶中强约35倍。根据模型,强烈的阻滞作用主要是由于与连续的单油酸甘油酯膜的流体动力学相互作用是比与离散的凝胶纤维的空间相互作用(碰撞)更强的摩擦源。第二个影响是规则的液晶比琼脂糖凝胶的随机网络具有更窄的孔径分布。与实验一致,这两种影响共同预测立方相中的阻滞作用比具有相同平均孔半径的琼脂糖凝胶中的阻滞作用强30倍。