PEG adenovirus purification - enough pure for retinal injection in rat?? (Feb/12/2009 )

I purify an adenovirus using PEG. We pretend to use it to test gene expression in retinal by injection. But I wonder if viruses are pure enough for this use?

Has anybody purify viruses that way and use them in animals???

aztecan princess on Feb 12 2009, 10:04 PM said:

I havn't purified virus that way. One thing can be to run a coomasie gel and you would only see the viral proteins and any thing else is contamination. Also you could inject it into the eye and see for any reaction in the eye, actually this is the best determinant of purity.

I'd worry about cellular proteins that come with it., which may cause more serious immune reaction than adv alone for your in vivo study.

You are right, we will check it in the animal, but I’m afraid that as you said we’ll have immune reaction problems… I just wanted to check if someone already has done.
Thank for your opinion!!

Definitely not pure enough.....its more of a concentrating procedure than a purification procedure. You could run the PEG concentrated virus on a sucrose or CsCl gradient to purify it

Preferably sucrose gradient, CsCl is very toxic to you and the rats.