A comparative study of different strategies for removal of endotoxins from bacteriophage preparations

作者：Van Belleghem Jonas D^*; Merabishvili Maya; Vergauwen Bjorn; Lavigne Rob; Vaneechoutte Mario

来源：Journal of Microbiological Methods, 2017, 132: 153-159.

DOI：10.1016/j.mimet.2016.11.020

• Endotoxins
• Lipopolysaccharides (LPS)
• Bacteriophages
• Pseudomonas aeruginosa
• Staphylococcus aureus
• Endotoxin removal
• Endotoxin quantification

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.

• 出版日期2017-1
• 单位KU Leuven