Table 1.

Advantages and Drawbacks of Extracellular Vesicle Isolation Methods

Isolation method	Advantages	Drawbacks
Differential ultracentrifugation	• Common method	• Low scalability
	• Medium to high yield	• Labor-intensive
		• Time-consuming
		• Possible aggregation or rupture of EVs
		• Possible contamination with lipoproteins
Density gradient ultracentrifugation	• Potential to identify EV subpopulations	• Low scalability
	• High purity	• Labor-intensive
	• Low amount of non-EV contaminants	• Time-consuming
Size-exclusion chromatography	• Fast and straightforward protocol	• Often needs another method to concentrate isolated EVs
	• Preserved integrity of isolated EVs
	• Highly scalable
Ultrafiltration	• Straightforward protocol	• Less common method
Tangential flow filtration	• Good EV yield	• Often needs another method to concentrate isolated EVs
	• No EV aggregation
	• Batch-to-batch consistency	• Favorable for functional EV studies
	• Suitable for large starting volumes
Immunoaffinity-based capture	• High purity	• Lower yield
	• Possibility of isolating subpopulations expressing a certain marker	• May not isolate all EVs
		• Higher cost
Precipitation	• Fast and straightforward protocol	• High amount of non-EV contaminants
		• Polymer remnants in the isolated EV sample