Limits...
Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials.

Merabishvili M, Pirnay JP, Verbeken G, Chanishvili N, Tediashvili M, Lashkhi N, Glonti T, Krylov V, Mast J, Van Parys L, Lavigne R, Volckaert G, Mattheus W, Verween G, De Corte P, Rose T, Jennes S, Zizi M, De Vos D, Vaneechoutte M - PLoS ONE (2009)

Bottom Line: 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.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, phiKMV and phage G1.The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.

View Article: PubMed Central - PubMed

Affiliation: Eliava Institute of Bacteriophage, Microbiology and Virology, Tbilisi, Georgia.

ABSTRACT
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 successive 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, phiKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.

Show MeSH

Related in: MedlinePlus

BFC-1 transmission electron micrographs.a) S. aureus bacteriophage ISP, a member of the Myoviridae family. Bar: 100 nm. b) PNM bacteriophages (Podoviridae) freed from a burst P. aeruginosa bacterium. Bar: 500 nm. c) Bacteriophage 14/1 attaching to the P. aeruginosa cell wall. Bar: 200 nm. d) ISP bacteriophages (Myoviridae) attached to S. aureus. Bar: 500 nm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2654153&req=5

pone-0004944-g004: BFC-1 transmission electron micrographs.a) S. aureus bacteriophage ISP, a member of the Myoviridae family. Bar: 100 nm. b) PNM bacteriophages (Podoviridae) freed from a burst P. aeruginosa bacterium. Bar: 500 nm. c) Bacteriophage 14/1 attaching to the P. aeruginosa cell wall. Bar: 200 nm. d) ISP bacteriophages (Myoviridae) attached to S. aureus. Bar: 500 nm.

Mentions: Microscopy confirmed that BFC-1 only contained bacteriophage particles with the expected morphology (Figure 4) and the expected target strain activity.


Quality-controlled small-scale production of a well-defined bacteriophage cocktail for use in human clinical trials.

Merabishvili M, Pirnay JP, Verbeken G, Chanishvili N, Tediashvili M, Lashkhi N, Glonti T, Krylov V, Mast J, Van Parys L, Lavigne R, Volckaert G, Mattheus W, Verween G, De Corte P, Rose T, Jennes S, Zizi M, De Vos D, Vaneechoutte M - PLoS ONE (2009)

BFC-1 transmission electron micrographs.a) S. aureus bacteriophage ISP, a member of the Myoviridae family. Bar: 100 nm. b) PNM bacteriophages (Podoviridae) freed from a burst P. aeruginosa bacterium. Bar: 500 nm. c) Bacteriophage 14/1 attaching to the P. aeruginosa cell wall. Bar: 200 nm. d) ISP bacteriophages (Myoviridae) attached to S. aureus. Bar: 500 nm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2654153&req=5

pone-0004944-g004: BFC-1 transmission electron micrographs.a) S. aureus bacteriophage ISP, a member of the Myoviridae family. Bar: 100 nm. b) PNM bacteriophages (Podoviridae) freed from a burst P. aeruginosa bacterium. Bar: 500 nm. c) Bacteriophage 14/1 attaching to the P. aeruginosa cell wall. Bar: 200 nm. d) ISP bacteriophages (Myoviridae) attached to S. aureus. Bar: 500 nm.
Mentions: Microscopy confirmed that BFC-1 only contained bacteriophage particles with the expected morphology (Figure 4) and the expected target strain activity.

Bottom Line: 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.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, phiKMV and phage G1.The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.

View Article: PubMed Central - PubMed

Affiliation: Eliava Institute of Bacteriophage, Microbiology and Virology, Tbilisi, Georgia.

ABSTRACT
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 successive 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, phiKMV and phage G1. The bacteriophage cocktail is currently being evaluated in a pilot clinical study cleared by a leading Medical Ethical Committee.

Show MeSH
Related in: MedlinePlus