I am trying to produce a large bulk of phages. The conventional methods are for low scale, ei, using chromatographic columns. Any suggestions?
Depending on your needs you may find tangential-flow ultrafiltration works well; we routinely use a 30kd ultrafiltration membrane, and can to purify phage with near 100 % efficiency from 200L of seawater. However, the efficiency can depend on the medium, and the specific conditions. More details can be found at: http://aem.asm.org/content/57/3/721.long
PEG (polyethelyene glycol) precipitation in the cold is the standard means of phage isolation. The precipitate can then be dialyzed into appropriate buffer. This can then be run in a CsCl2 equilibrium density gradient to generate high titer ultra pure phage.
This correct reference for the PEG and CsCl2 procedures should have been our prior 1978 article, now attached,
Determination of capsid size by satellite bacteriophage P4,
Proc Natl Acad Sci USA, Shore, Deho, Tsipis & Goldstein, 1978
Article Transcriptional control of capsid size in the P2:P4 bacterio...
Recently, fast and efficient chromatographic methods have also been developed, which can be scaled up successfully. As an example see:
http://www.biaseparations.com/applications/viruses-vlps/product/24-single-step-method-for-purification-of-bacteriophage-vdx-10-on-cim-qa-disk-monolithic-column-and-method-scale-up
Regard
Luca A. V.
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