Exosomes, which are freely available in plasma and other body fluids, have attracted a great deal of interest as therapeutic and diagnostic biomarkers. With years of experiences in exosome technology, Creative Biolabs has developed validated protocols to purify exosomes from both plasma and other body fluids with decreased co-purification of non-exosomal proteins.
To enhance the concentration and quality of exosome, many techniques have been developed for exosome purification, including ultracentrifugation and microfiltration, size exclusion chromatography (SEC), and immunoaffinity capture.
Ultracentrifugation is generally considered to be the gold standard method for extracellular vesicles isolation. This technique separates vesicles from other sample components depending on their sedimentation rates that are mostly determined by size and density. Microfiltration is an isolation/enrichment method that utilized hydrophilized polyvinylidene difluoride membrane to easily purify exosomes from samples with decreased co-purification of abundant non-exosomal proteins.
Fig.1 Ultracentrifuge.
SEC allows to separate sample components based on the differential rate of movement through a gel matrix of different sized components. The smaller a component, the more it is able to penetrate into the gel matrix, the longer it retains in the column and the later the elution. In the context of exosome isolation, the vesicles can be separated from smaller protein and nucleic acid contaminants.
Immunoaffinity capture methods use antibody-coated beads to selectively isolate vesicles displaying a surface marker of interest. Preconcentrated extracellular vesicle samples are incubated with magnetic beads coated with an antibody against the target marker, allowing the specific pulldown of extracellular vesicles expressing target on their surface.
Enhanced purification of exosomes benefits downstream analyses, including more accurate quantification of vesicles and characterization. Creative Biolabs provides the high-purity and high-yield exosomes for global clients. We also believe that our service will make your project achieve twice the result with half the effort. If you have any demands in exosome preparation, please don’t hesitate to contact us for more information.
A: Exosome purification is to obtain high-quality exosome samples for further study of their composition and function. Purification can remove other cellular components and impurities, making the study of exosomes more precise.
A: Exosome purification methods include ultracentrifugation, density gradient centrifugation, size exclusion chromatography, affinity chromatography, etc. Choosing an appropriate method depends on the nature of the sample and the purpose of the study.
A: Purity can be assessed by observing exosome morphology with electron microscopy, detecting specific proteins by Western blot, and nanoparticle tracking analysis (NTA).
A: Exosomes should generally be stored in a frozen state at -80°C or in liquid nitrogen tanks to ensure their stability and maintain their functionality.