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Purification of virus particles


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#1 lene

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Posted 24 August 2009 - 11:41 PM

Hello!

For particle size analysis of virions I have to purify the samples and get rid of the sucrose, which was added for the purification step with gradient ultracentrifugation.
I tried different systems like dialysis, filtration, tca-precipitation, ... . The problem is that the virus particles are very sticky and sticks to the membranes, the walls of the tubes, etc. so that in the end after purification no virus is left. I started adding tween 20 for keeping the protein soluble, but it does not help that much.

Has anybody an idea how to purify the virus sample without loosing all the material? Some different inexpensive methods? And in addition the virus particles should stay in their native state.

Thanks, lene

#2 molgen

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Posted 03 September 2009 - 01:57 AM

In the lab next door to mine during my masters they had a similar problem.
They ended up doing immunoprecipitation of the particles.
They did it as the last resort but managed to get by.
The yield was not that good but it was better then nothing

#3 lene

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Posted 07 September 2009 - 06:56 AM

Hello,
thanks for the hint! Do you know what they did in more detail?
What they used (magnetic beads, agarose beads)? How they broke up the linkage between the virus and the antibody without destroying the virus?




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