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We present an alternative bioprocess for bacteriophages involving room temperature coprecipitation of an aqueous mixture of phage (Siphoviridae) and a crystallizable carrier (glutamine or glycine) in excess of water miscible organic solvent (isopropanol or isobutanol). The resultant suspension of phage-coated microcrystals can be harvested by filtration and the residual solvent removed rapidly by air-drying at a relative humidity of 75%. Albumin or trehalose added at 5% w/w of the crystalline carrier provide for better stabilization of the phage during co-precipitation. Free-flowing dry powders generated from an aqueous solution of phage (similar to 13 log10 pfu/mL) can be reconstituted in the same aqueous volume to a phage titer of almost 10 log10 pfu/mL; high enough to permit subsequent formulation steps following bioprocessing. The phage-coated microcrystals remain partially stable at room temperature for at least one month, which compares favorably with phage immobilized into polyester microcarriers or lyophilized with excipient (15% polyethylene glycol 6000 or 0.10.5 M sucrose). We anticipate that this bioprocessing technique will have application to other phage families as required for the development of phage therapies. (c) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012
|Number of pages||9|
|Publication status||Published - Apr 2012|
- Staphylococcus aureus
- protein-coated microcrystals
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- 1 Finished
New Approaches to High Throughput Protein, Isolation, Purification and Concentration
Moore, B., Cormack, P., Sefcik, J. & Van Der Walle, C.
BBSRC (Biotech & Biological Sciences Research Council)
29/10/07 → 28/06/11