Extracellular Vesicles (EVs) are small lipid-bound vesicles secreted by cells into the extracellular space, which can carry various biomolecules such as proteins, lipids, DNA, RNA, and metabolites. These biomolecules can reflect the physiological and pathological status of the cells of origin, and can also modulate the functions of recipient cells. Therefore, studying the content of EVs can provide valuable insights into the mechanisms and roles of EV-mediated intercellular communication, as well as potential applications in biomarker discovery and drug delivery.

However, before studying the content of EVs, one needs to disrupt the lipid bilayer of the EVs to release the biomolecules from the vesicles. This can be done by using various methods, such as physical, chemical, or enzymatic methods. These methods usually involve breaking the lipid bilayer of the EVs using detergents, chaotropic agents, solvents, enzymes, electrical current, or sonication. The choice of method depends on the type and quality of biomolecules required, the type and amount of EVs available, and the downstream analysis planned.

In this blog post, I will explain some of the common methods for disrupting EV membrane and their advantages and disadvantages. I will also provide some links for further information and resources.

Physical methods

Physical methods involve applying physical force to the EVs to break their lipid bilayer and release their biomolecules. Physical methods can be performed using various devices, such as nitrogen cavitation¹, extrusion via porous membrane², or sonication³. These devices usually generate high pressure or shear force that disrupts the EV membrane and reassembles them into smaller vesicles containing biomolecules.

Some advantages of physical methods are:

• They are fast and efficient
• They do not require additional reagents or chemicals
• They can be applied to various types of EVs and biomolecules

Some disadvantages of physical methods are:

• They can cause heat generation, foaming, or aggregation of EVs
• They can cause damage or degradation of biomolecules
• They can introduce contamination or artifacts from the device

Chemical methods

Chemical methods involve using detergents, chaotropic agents, or solvents to disrupt the EV membrane and solubilize their biomolecules. Chemical methods can be performed using various reagents, such as sodium dodecyl sulfate (SDS)⁴, guanidine thiocyanate, phenol-chloroform, or Trizol. These reagents usually interact with the lipid molecules of the EV membrane and disrupt their structure and function.

Some advantages of chemical methods are:

• They are effective and gentle
• They can preserve the integrity and activity of biomolecules
• They can be combined with other methods for further purification

Some disadvantages of chemical methods are:

• They require careful handling of hazardous chemicals
• They can cause denaturation, coagulation, or contamination of biomolecules
• They can interfere with some downstream analysis methods

Conclusion

Disrupting EV membrane is a necessary step in studying the content of EVs, as it helps to release the biomolecules from the vesicles and make them accessible for further analysis. Disrupting EV membrane can be done by using various methods, such as physical, chemical, or enzymatic methods. These methods have different advantages and disadvantages, depending on the type and quality of biomolecules required, the type and amount of EVs available, and the downstream analysis planned. By following these steps and tips, you can disrupt EV membrane effectively and efficiently, and study their content.