We would like to do exosomes extraction from brain homogenates but we do not have ultracentrifuge. I was wondering if someone tried any alternative protocol that do not need an ultracentrifuge.
Hello Dr. Sara Elfarrash
Although ultracentrifugation is the most widely used exosome purification protocol and has long been considered the gold standard for isolation of relatively homogenous size populations of exosomes, you could use a new exosome isolation method, termed as rinsing separation method which the investigators in the attached paper below have developed, and they have compared its advantages and weaknesses to the existing ultracentrifugation and ExoQuick precipitation methods.
They claim that this new method was particularly useful for protein analyses due to the reduced amount of non-exosome protein contamination. Additionally, this method was more time-efficient and cost-efficient compared to ultracentrifugation.
https://www.europeanreview.org/wp/wp-content/uploads/5074-5083.pdf
However, a major limitation of this study is that the exosomes used were isolated from only one cell type. Although each step was optimized, there is a possibility that other cell types may require modified parameters to successfully isolate highly pure exosomes. Therefore, rinsing separation method should have to be optimized for use in a variety of cell types.
Hope this is helpful!
Best.
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