IMMUNOMAGNETIC SEQUENTIAL ULTRAFILTRATION PLATFORM FOR ENRICHMENT AND PURIFICATION OF EXTRACELLULAR VESICLES FROM BIOFLUIDS

摘要

Methods for purifying and isolating extracellular vesicles (EVs) from a biofluid using a sequential processing. Tangential flow filtration is applied to the biofluid to increase the concentration of EVs in the biofluid. After this is achieved, enrichment mode is halted and the biofluid is processed in diafiltration mode to remove contaminants (up to 99.9%). After performing the tangential flow filtration step, the concentration of EVs in the biofluid is further increased by ultracentrifugal filtration. After performing the ultracentrifugal filtration step, EVs of a particular target type are separated from other EVs by immunomagnetic affinity separation. In some implementations, the methods are used to isolate and quantify tumor EVs for cancer evaluation. Additionally, these methods can be used with a scaling factor to quantify EVs from a less concentrated biofluid such as, for example, urine.