Asked 12th Aug, 2014

Does anybody know a good method for large scale bacteriophage purification?

I am trying to produce a large bulk of phages. The conventional methods are for low scale, ei, using chromatographic columns. Any suggestions?

Most recent answer

5th Sep, 2014
Luca A Vitali
University of Camerino
Recently, fast and efficient chromatographic methods have also been developed, which can be scaled up successfully. As an example see:
Luca A. V.
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All Answers (4)

13th Aug, 2014
Curtis A Suttle
University of British Columbia - Vancouver
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: 
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13th Aug, 2014
Rene Kaden
Uppsala University
If you want to extract almost pure phages from sediments I woud suggest to use 10% beef extract (Difco; 1 ml/g sediment) followed by 1 h shaking at room temperature and 20 min centrifugation at 3000 g. The supernatant contains the extracted phages. There exist some protocols for enrichment: e.g. ISO 10705-1:1995 for F-specific ribonucleic acid bacteriophages
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4th Sep, 2014
Richard Goldstein
Boston University
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

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SDS-PAGE analysis of the phage particle proteins. CsCl gradient purified tf preparation was used. Putative gene products (gp) shown were identified on the basis of molecular weight estimations. (TIF)
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