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Bioprocessing of bacteriophages via rapid drying onto microcrystals

Moore, Barry and Alvarez-Gonzalez, Eva and Alfadhel, Munerah and Mane, Parag and Ford, Steven J. and van der Walle, Christopher F. (2012) Bioprocessing of bacteriophages via rapid drying onto microcrystals. Biotechnology Progress, 28 (2). pp. 540-548. ISSN 8756-7938

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Abstract

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