TY - JOUR
T1 - Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials
AU - Merabishvili, Maya
AU - Pirnay, Jean Paul
AU - Verbeken, Gilbert
AU - Chanishvili, Nina
AU - Tediashvili, Marina
AU - Lashkhi, Nino
AU - Glonti, Thea
AU - Krylov, Victor
AU - Mast, Jan
AU - Van Parys, Luc
AU - Lavigne, Rob
AU - Volckaert, Guido
AU - Mattheus, Wesley
AU - Verween, Gunther
AU - De Corte, Peter
AU - Rose, Thomas
AU - Jennes, Serge
AU - Zizi, Martin
AU - De Vos, Daniel
AU - Vaneechoutte, Mario
PY - 2009/3/20
Y1 - 2009/3/20
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=64049105821&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0004944
DO - 10.1371/journal.pone.0004944
M3 - Article
C2 - 19300511
AN - SCOPUS:64049105821
SN - 1932-6203
VL - 4
JO - PLoS ONE
JF - PLoS ONE
IS - 3
M1 - e4944
ER -