I am looking to isolate exosomes from human plasma samples by a magnetic bead-based method. Is it recommended that I filter the plasma samples through a 0.22 micron cellulose acetate filter before exosome isolation? Will this contribute to a loss in exosomes being isolated?
Hello Harshini Sriram
Yes, you can filter human plasma through 0.22 micron cellulose acetate filter. It should not cause a loss in exosomes.
The filter used for filtration of biological samples should be wettable with no charge and negligible or completely not binding to biological components like proteins, RNA, DNA, polysaccharides, lipids and other biological entity. The type of membrane material having such properties are polyether sulfone or cellulose acetate. So, you can go ahead and filter plasma samples through 0.22micron cellulose acetate filter. Please prewet the membrane filter before use to avoid loss of exosomes.
I hope this is helpful.
Best Wishes.
Hi Harshini Sriram
I am curious to know: are you using a commercial kit? If so is there any particular one which has been successful for you?
Best,
Harini
Hi Harini Lakshminarayanan
Yes, we are using a commercial kit. FUJIFILM Wako Pure Chemical Corporation offers a number of exosome isolation kits for culture supernatants and/or serum and plasma. These are what we are using as of now.
We have, in the past, also tried the Total exosome isolation reagent from Thermofisher and the miRCURY exosome kit from Qiagen.
Do let me know if you'd have any suggestions/opinions for us on this front.
Best wishes.
I have used 2 commercial kits but for me none has worked fine. They are isolating vesicles which are not giving any signal of exosomal markers on flow cytometry.
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