不对称流场-流动分馏(AF4)作为一种基于尺寸的分离技术引起了相当大的关注,由于其温和的分离条件,宽工作范围(分子量约为103至109Da或粒径为1nm至1μm),和多功能性。AF4主要用于测量颗粒大小,多分散性,和各种系统的物理稳定性,如(生物)大分子和纳米颗粒。与尺寸排阻色谱(填充柱)相比,AF4(开放通道)允许分离,同时保留不稳定结构。监测不同化合物之间和非常复杂的基质中的相互作用是可能的。结构的保存以及结构特征与活性和功能性的相关性可以支持疾病的新治疗策略和具有改进特性的新材料的开发。在这次审查中,详细概述了AF4在各种系统之间相互作用研究方面的发展,如蛋白质-蛋白质,聚合物-聚合物,纳米颗粒药物,和纳米颗粒蛋白。AF4和其他不太常用的FFF类型的前景和障碍,用于研究复杂和脆弱系统内的相互作用。将AF4耦合到各种检测系统可以极大地有助于理解相互作用/关联过程并提供有关相互作用动力学的信息。本审查旨在提供有关信息类型的全面文档(结构,形态学,化学)关于AF4可以检索的分子相互作用。
Asymmetrical flow field-flow fractionation (AF4) has attracted considerable attention as a size-based separation technique, due to its mild separation conditions, broad working range (from approximately 103 to 109 Da molecular mass or from 1 nm to 1 μm particle diameter), and versatility. AF4 is primarily being used to measure particle size, polydispersity, and physical stability of various systems, such as (bio)-macromolecules and nanoparticles. In comparison with size-exclusion chromatography (packed column), AF4 (open channel) allows separation while preserving labile structures. Monitoring of interactions between different compounds and in very complex matrices is possible. Preservation of the structure and correlation of structural characteristics with activity and functionality can bolster the development of new therapeutic strategies for diseases and new materials with improved properties. In this
review, a detailed overview is presented of developments in AF4 for interaction studies between various systems, such as protein-protein, polymer-polymer, nanoparticle-drug, and nanoparticle-protein. The prospects and obstacles for AF4, and other less-commonly used types of FFF, for studying interactions within complex and fragile systems are covered. Coupling AF4 to a variety of detection systems can greatly contribute to the understanding of the interaction/association processes and provide information on the interaction kinetics. This
review is intended to provide comprehensive documentation on the types of information (structural, morphological, chemical) on molecular interactions that can be retrieved by AF4.