Someone who has tried some ultracentrifugation-free AAV viral vector purification protocol could recommend me a protocol to obtain high titers and purity of AAVs. I would really appreciate any help.
Greetings,
I hope this may help you:
https://www.addgene.org/protocols/aav-purification-iodixanol-gradient-ultracentrifugation/
Regards
Oh I see, my mistake, didn't get that right. Well, if there is no chance to get an ultracentrifuge, you can always run cessium chloride gradient at x5000 g for 24 hours, using 50 mL falcon tubes. Another method that you can use to isolate AAV is size exclusion chromatography, there are several columns that reach to 1-8 MDa. AAV capsid must be around 2.7 or more MDa, like other parvovirus capsids. I've used SEC as a first step of purification for PstDV1, followed by ultracentrifugation gradient, however, you could get your AAV's particles with enough purity after 2-4 SEC runs. Next, you can find two protocols for parvoviral VLPs that you can easily adjust to your experimental conditions.
Good luck.
I found this paper which I will try.
- PMC3374034
- Rapid and simplified purification of recombinant adeno-associated virus. J Virol Methods. 2012 Aug; 183(2): 139–146.
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