Institute of Bioengineering (IBI) and Institute of Chemical Sciences and Engineering (ISIC), Ecole Polytechnique Fédérale de Lausanne, Lausanne CH-1015, Switzerland.
Methods. 2011 May;54(1):92-100. doi: 10.1016/j.ymeth.2010.12.003. Epub 2010 Dec 10.
Analytical ultracentrifugation (AUC) primarily serves to investigate hydrodynamic and thermodynamic properties of natural and synthetic macromolecules and colloids in solution, dispersion or suspension. Beside such more conventional use, AUC can support materials development particularly by combining different optical systems, if the AUC is equipped with such, or using complementary data evaluation approaches. In this context, an Optima XL-I equipped with absorbance (AO) and interference optics (IO) was used alone or complementary to study the success of conjugation of biopolymers, to evaluate the completeness of the incorporation of macromolecules into micelles and vesicles, and to analyze the composition and homogeneity of macromolecular assemblies. The combination of AO and IO proved covalent binding of concanavalin A to dextran without macromolecular degradation as well as the formation of mixed micelles composed of two types of block copolymers. Further, AUC contributed to analyze the homogeneity, purity, size and size distribution of carbon monoxide-releasing macromolecular assemblies. These case studies revealed that the application possibilities of AUC are by far not completely discovered but can still be extended.
分析超速离心(AUC)主要用于研究天然和合成大分子及胶体在溶液、分散体或悬浮液中的流体力学和热力学性质。除了这种更传统的用途之外,AUC 还可以通过结合不同的光学系统来支持材料开发,如果 AUC 配备了这种系统,或者使用补充的数据评估方法。在这种情况下,单独使用或与互补的吸收(AO)和干涉光学(IO)一起使用 Optima XL-I 来研究生物聚合物缀合的成功、评估大分子完全掺入胶束和囊泡的程度,以及分析大分子组装的组成和均一性。AO 和 IO 的组合证明了伴刀豆球蛋白 A 与葡聚糖的共价结合,而没有大分子降解,以及由两种类型的嵌段共聚物组成的混合胶束的形成。此外,AUC 有助于分析一氧化碳释放大分子组装的均一性、纯度、大小和大小分布。这些案例研究表明,AUC 的应用可能性远未完全被发现,但仍可以进一步扩展。