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We describe the small-scale, laboratory-based, production and quality control of a cocktail, consisting of exclusively lytic bacteriophages, designed for the treatment of  Pseudomonas aeruginosa  and  Staphylococcus aureus  infections in burn wound patients. Based on succesive selection rounds three bacteriophages were retained from an initial pool of 82  P. aeruginosa  and 8  S. aureus  bacteriophages, specific for prevalent  P. aeruginosa  and  S. aureus  strains in the Burn Centre of the Queen Astrid Military Hospital in Brussels, Belgium. This cocktail, consisting of  P. aeruginosa  phages 14/1 ( Myoviridae ) and PNM ( Podoviridae ) and  S. aureus  phage ISP ( Myoviridae ) was produced and purified of endotoxin. Quality control included Stability (shelf life), determination of pyrogenicity, sterility and cytotoxicity, confirmation of the absence of temperate bacteriophages and transmission electron microscopy-based confirmation of the presence of the expected virion morphologic particles as well as of their specific interaction with the target bacteria. Bacteriophage genome and proteome analysis confirmed the lytic nature of the bacteriophages, the absence of toxin-coding genes and showed that the selected phages 14/1, PNM and ISP are close relatives of respectively F8, φKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.

Quality-Controlled Small-Scale Production of a Well-Defined Bacteriophage Cocktail for Use in Human Clinical Trials