[RASMB] detecting virus particles with AUC

[John Philo]

Veysel,

Yes, this can be done, see:
Berkowitz, S. A. and Philo, J. S. (2007). Monitoring the homogeneity of
adenovirus preparations (a gene therapy delivery system) using analytical
ultracentrifugation.  Anal. Biochem. 362, 16-37. 

The speed will of course depend on the virus monomer size but also on the
extent of aggregation. Some adenovirus preparations were so aggregated (like
1,000-10,000-mers) it was difficult to capture everything at 3,000 rpm
(that's about as slow as the instrument wants to go). I've run adenovirus
usually at 3,000 rpm and AAV (a smaller one) at around 10,000 rpm, so that
is probably the right ballpark, but you'll probably need to do a preliminary
run to get the right speed.

Hopefully this virus isn't a serious biosafety issue, but you will want to
think about the fact that if your cell leaks the vacuum pump will
potentially vaporize virus into the room. Also some viruses may require
detoxification with bleach, but that will destroy your aluminum housings.

John

-----Original Message-----
From: rasmb-bounces at rasmb.bbri.org [mailto:rasmb-bounces at rasmb.bbri.org] On
Behalf Of Veysel Kayser
Sent: Thursday, September 25, 2008 10:34 AM
To: rasmb at server1.bbri.org
Subject: [RASMB] detecting virus particles with AUC

Dear AUC experts,

Is it possible to detect virus particles (aggregated and disassembled
viruses as well monomers) in AUC? If so, what would be the recommended
concentration and speed for spinning them? In addition, what is the
sensitivity of AUC, e.g. can one detect monomer, dimer and smaller particles
at the same time? The virus has a bullet shape with 100nm X 200nm size. Any
input would be appreciated.

Veysel
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