title: Preparation of endotoxin-free bacteriophages.
abstract: Bacteriophages (phages) are bacterial viruses that interact with bacterial walls and invade bacterial cells. Moreover, they disturb bacterial metabolism and lead to bacteria lysis. In the case of Gram-negative bacteria crude phage cultures, apart from the phages themselves, the bacterial debris, bacterial proteins and nucleic acids contain endotoxins. These endotoxins (lipopolysaccharides) posses a high degree of toxicity in vitro and in vivo, and their removal is essential for safety in antibacterial bacteriophage therapy. An effective, scaleable purification of bacteriophages from endotoxins was accomplished by sequential ultrafiltration through polysulfone membrane (30 nm) followed by chromatography on sepharose 4B and Matrex Cellulofine Sulfate. The phage fraction after gel filtration chromatography routinely contained endotoxins in the 150-2500 EU/ml range. The procedure yielded bacteriophages contaminated with as little as 0.4-7 EU/ml (Limulus assay). This value lies within the permitted level for intravenous applications (5 EU/kg/h by European Pharmacopoeia, 1997).
authors.affiliation: Laboratory of Biomedical Chemistry, Department of Experimental Oncology, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wrocław, Poland. borat@iitd.pan.wroc.pl
authors.forename: Janusz
authors.initials: J
authors.lastname: Boratyński
authors.forename: Danuta
authors.initials: D
authors.lastname: Syper
authors.forename: Beata
authors.initials: B
authors.lastname: Weber-Dabrowska
authors.forename: Marzanna
authors.initials: M
authors.lastname: Łusiak-Szelachowska
authors.forename: Gryzelda
authors.initials: G
authors.lastname: Poźniak
authors.forename: Andrzej
authors.initials: A
authors.lastname: Górski
mesh_terms: D000818:Animals; D001435:Bacteriophages / Q000737:chemistry* / Q000378:metabolism*; D004731:Endotoxins / Q000302:isolation & purification*; D004926:Escherichia coli / Q000821:virology; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000821:virology
chemical_list: D004731:Endotoxins