Hydrophobic Chromatography for Extracellular Vesicle Isolation

The patent changes the separation parameter from physical properties (size, density) to chemical property (hydrophobicity). By using hydrophobic interaction chromatography with a stationary phase containing hydrophobic functional groups, the method exploits differences in hydrophobicity between extracellular vesicles and lipoproteins to achieve selective separation, thereby eliminating lipoprotein contamination while maintaining high purity of isolated extracellular vesicles

The patent introduces a hydrophobic stationary phase as an intermediary medium for separation. This stationary phase with specific hydrophobic functional groups acts as a mediator that selectively interacts with extracellular vesicles through hydrophobic forces, enabling their separation from lipoproteins without direct comparison of their similar physical properties

The patent replaces the complex mechanical ultracentrifugation system with a simpler chromatography system based on hydrophobic interaction. Instead of using high-speed rotation and density gradient mechanisms, the method uses a stationary phase with hydrophobic functional groups that selectively retain extracellular vesicles, allowing separation to occur under much milder and simpler conditions with faster processing time

The patent changes the separation mechanism from mechanical force-based (centrifugal force in ultracentrifugation) to chemical interaction-based (hydrophobic interaction). This parameter change enables the use of simpler equipment without high-speed centrifuges while achieving faster isolation rates and reduced procedural complexity