During viral propagation enriched medium contains BSA, which is tightly associated with virus. I cannot seem to remove this even by purification by SEC. I need virus void of BSA association. How can we eliminate this problem? Can anybody help?
When analysed SEC fraction on ELISA with anti-BSA and viral specific mab we can see both.
I agree with Vladimir Loparev.You can ultracentrifuge using sucrose or CsCl gradient.
A little more information will be helpful. Are you working with an enveloped virus or a naked virus. This is important as CsCl gradients will, for the most part, de-envelope virus particles thus making them non-infectious, however, they work great for naked viruses. If enveloped, you may want to use a sucrose-density gradient ultracentrifugation method to isolate/purify your virus...as long as the bouyant density of virions, in sucrose, has been characterized.
We routinely purify cell culture derived retroviruses from media containing 10% FBS. I agree that sucrose density gradient centrifugation is a good method. We also simply pellet virus through 20% sucrose pads - which is much easier than the density gradient method. Both methods eliminate most (but not all) of the BSA. A further removal of BSA can be accomplished by resuspending the purified virus in a serum-free buffer and direct pelleting. We do this using a Beckman TLA100.3 rotor because it uses small polyallomer tubes which do not adsorb the virus. Using larger, non-polyallomer tubes may result in significant loss of virus. Please feel free to contact me if you would like to discuss this in detail. Good luck!
I agree with Joshua Geltz. Information about enveloped and non-enveloped viruses is important.
I agree with Joshua Geltz. The sucrose density gradient ultracentrifugation is a peferred method for virus puriffication about enveloped viruses.
I'm also interested in this issue. Does anybody know what kind of interactions are those between BSA-viral particles? I see you all are talking about density gradients, but, would an ionic exchange chromatography work? Do those chromatographies work in both enveloped and non-enveloped virus?
Thak you all!
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