How to concentrate retrovirus produced by Phoenix-Ampho cell - (Nov/10/2008 )
hi,
I recently got the retrovirus produced by Phoenix-ampho system, but the titer was too low, so want to concentrate the virus solution. But I was told it couldn't be done by untracentrifugation because it would lose some proteins on the surface of retrovirus. Who knows how to concentrate this virus? I guess low speed centrifugation should work, but I need to choose a suitable MWCO membrane. Who knows which MWCO membrane can cut off the retrovirus.
-rabbitwrong-
QUOTE (rabbitwrong @ Nov 10 2008, 10:14 AM)
hi,
I recently got the retrovirus produced by Phoenix-ampho system, but the titer was too low, so want to concentrate the virus solution. But I was told it couldn't be done by untracentrifugation because it would lose some proteins on the surface of retrovirus. Who knows how to concentrate this virus? I guess low speed centrifugation should work, but I need to choose a suitable MWCO membrane. Who knows which MWCO membrane can cut off the retrovirus.
I recently got the retrovirus produced by Phoenix-ampho system, but the titer was too low, so want to concentrate the virus solution. But I was told it couldn't be done by untracentrifugation because it would lose some proteins on the surface of retrovirus. Who knows how to concentrate this virus? I guess low speed centrifugation should work, but I need to choose a suitable MWCO membrane. Who knows which MWCO membrane can cut off the retrovirus.
Use PEG. See the AquaRNA Viral Protocol for details.
-chessplayer-
There are several papers regarding this, too. We spin at 20Kg, 4h, 4oC.
-Almasy-
It is for phoenix-ampho? I know adenovirus can be concentrated by this? But for retrovirus, centrifuging will lose some important proteins.Just centrifuge it? Don't use the membrane to cut the molecule weight?
QUOTE (Almasy @ Nov 12 2008, 12:00 AM)
There are several papers regarding this, too. We spin at 20Kg, 4h, 4oC.
-rabbitwrong-
QUOTE (rabbitwrong @ Nov 14 2008, 10:44 AM)
It is for phoenix-ampho? I know adenovirus can be concentrated by this? But for retrovirus, centrifuging will lose some important proteins.Just centrifuge it? Don't use the membrane to cut the molecule weight?
QUOTE (Almasy @ Nov 12 2008, 12:00 AM)
There are several papers regarding this, too. We spin at 20Kg, 4h, 4oC.
Yes, for retrovirus-phoenix cell. I filter the medium first to get rid of cell debris, then spin. Then resuspend the pellet in some ml of medium (volume as desired). You check the paper. They have done the tests and tell you what speed and how long can you spin. PubMed for retrovirus and concentrate, I think.
-Almasy-