Table 1.

Summary of the commonly used EV isolation methods.

Technique	Principle	Advantages	Disadvantages
Ultracentrifugation	Particles with different sizes and densities have different sedimentation rates during ultracentrifugation	Ease of use High purity Suitable for large volume preparation	Extremely tedious Time-consuming Low recovery High equipment cost Possible structure damage
Size exclusion chromatography	EVs pass through a porous stationary phase in which small particles enter into the pores resulting in the late elution	Maintain the native state of EVs High purity	Results in large volume of eluted samples
Ultrafiltration	EVs pass through a membrane with defined pore size or molecular weight cut-off	Fast isolation process Low equipment cost	Vesicle clogging and trapping
Immunoaffinity	Based on specific binding between surface marker proteins of EVs and immobilized antibodies	High purity and selectivity	High-cost antibodies Elution may damage native EV structure
Precipitation	Polymers decrease the solubility of EVs by creating the hydrophobic micro-environment	Ease of use High yield	Low purity Polymers affect downstream MS analysis