Suitable Agarose Beads for Extracellular Vesicle Chromatography

University of Toronto
Chemical/Biomedical Engineering
PhD Candidate

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Company:

Sigma-Aldrich

Product Name:

Sepharose® 4B

Catalog Number:

4B200-100ML

Extracellular vesicles (EVs) were isolated from conditioned cell culture media from various cell types via chromatography in columns created using sepharose 4B beads packed into a syringe with a stopper. Fractions were collected to analyze EV yield.

Experimental Design and Results Summary

Application

Size exclusion chromatography isolation of EVs

Starting Material

Conditioned media from various cell cultures

Tips

Wash through 1 column worth of PBS after packing and before adding sample/collecting fractions.

Results Summary

EVs were successfully isolated and concentrated in a subset of fractions, with other fractions containing separated protein aggregates. Nanoparticle tracking and the BCA protein assay was used to quantify particle concentration versus the amount of co-isolated protein in each fraction to ensure sufficient separation.

DOI or PMID #

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Summary

The Good

Relatively affordable way to make simple chromatography columns that can cost much more when purchased commercially

The Bad

Difficult to setup and time consuming to run, requires optimization and experience. Slightly lower yield than other techiques.

The Bottom Line

These beads provide a good resource for making in-house chromatography columns. Best if long term use makes optimization worthwhile. If not, then perhaps a commercial product is better for one-time use.

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