Evolving utility of exosomes in pancreatic cancer management

Table 5 Comparison of various isolation methods for exosomes

Conventional isolation of exosomes
Methods	Advantages	Disadvantages	Clinical use	Ref.
Ultracentrifugation	Widely used; high purity; protein and RNA components are not affected	Highly labour intensive; time-consuming; yields are typically low extensive training of personnel needed; expensive; inappropriate for the extraction of exosomes from a small amount of serum samples	Functional study of exosomes	[65,66]
Ultrafiltration	High yield; simple; less time-consuming; do not require the use of special equipment	Low purity, clogging of pores	Study of sample concentration; used in combination with other methods	[67]
Precipitation	Widely used; economical	Co-isolation of non-EV particles	For studies with very low purity requirements that do not require omics studies	[68]
Size exclusion chromatography, OR, and gel filtration	Fast, reliable, and inexpensive; maintain the biological activity and integrity of exosomes; high purity	Nanoscale contaminants like lipoproteins; extensive laboratory equipment requirements	Suitable for exosome research in those requiring high purity, omics, and large volume samples	[69]
Immunoaffinity capture	Convenient; not affected by exosome size; no need for expensive instruments	Expensive; low capacity; low yields	Suitable for the Separation of specific exosome subgroups	[70]
Emerging isolation methods
Stirred ultrafltration	Do not rely on equipment; less time consuming; reduces the destruction of exosomes during the process	Moderate purity of isolated exosomes; loss of exosomes during the process	Isolating exosomes from culture supernatant of bone marrow mesenchymal stem cells	[71]
ExoTIC (exosome total isolation chip)	Simple, easy-to-use, modular, and facilitates high-yield and high-purity EV isolation from biofluids	Special equipment requirements; lack of tests on clinical samples	Efficiently isolate EVs from small sample volumes; EV-based clinical testing from fingerprick quantities (10-100 μL) of blood	[72,73]
3D ZnO Nanoarrays	Multifunction; high sensitivity; downstream analysis is possible; enhance the capture of exosomes at a high flow rate	Relatively expensive	Widely used in biosensing and analysis aspects, powerful tools for effective purification and molecular analysis of exosome	[74,75]
Nano plasmon-enhanced scattering	Rapid, high-throughput, sensitive, and specifc method for the detection of exosomes from trace samples depending on the amount of scatter area, based on calculation of the proportion of the area that contains scattered light	High reagent cost; complex statistical tools; low capacity	Uses antibodies against the cellular markers CD81, CD63, and CD9, which are enriched on most exosome membranes	[76]

SEC: Size exclusion chromatography; EV: Extracellular vesicle.