期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 132, 期 -, 页码 153-159出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2016.11.020
关键词
Endotoxins; Lipopolysaccharides (LPS); Bacteriophages; Pseudomonas aeruginosa; Staphylococcus aureus; Endotoxin removal; Endotoxin quantification
资金
- FWO Vlaanderen [W0.016.14N]
- Institute for innovation through science and technology (IWT, Flanders, Belgium)
Bacterial endotoxins have high immunogenicity. Phage biology studies as well as therapeutic phage applications necessitate highly purified phage particles. In this study, we compared combinations of seven different endotoxin removal strategies and validated their endotoxin removal efficacy for five different phages (i.e. four Pseudomonas aeruginosa phages and one Staphylococcus aureus phage). These purification strategies included Endotrap HD column purification and/or CsCI density centrifugation in combination with Endotrap purification, followed by organic solvent (1-octanol), detergent (Triton X-100), enzymatic inactivation of the endotoxin using alkaline phosphatase and CIM monolytic anion exchange chromatography. We show that CsCI density purification of the P. aeruginosa phages, at an initial concentration of 10(12)-10(13) pfu/ml, led to the strongest reduction of endotoxins, with an endotoxin removal efficacy of up to 99%, whereas additional purification methods did not result in a complete removal of endotoxins from the phage preparations and only yielded an additional endotoxin removal efficacy of 23 to 99%, sometimes accompanied with strong losses in phage titer. (C) 2016 Elsevier B.V. All rights reserved.
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