ISSN Print 2713-2757
ISSN Online 2713-2765
SCIENTIFIC AND PRACTICAL REVIEWED JOURNAL OF FMBA OF RUSSIA
ORIGINAL RESEARCH
Comparison of methods for purification of bacteriophage lysates of gram-negative bacteria for personalized therapy
About authors
1 Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russia
2 Federal Research Center of Biotechnology, Moscow, Russia
Correspondence should be addressed: Roman B. Gorodnichev
Malaya Pirogovskaya, 1a, Moscow, 119435; moc.liamg@b.r.vehcindorog
About paper
Funding: all study expenses were covered by the funds allocated for the State Assignment on the Development of a personalized approach to the therapy of infections using virulent bacteriophages (Code: Bacteriophage).
Author contribution: Gorodnichev RB — planned the study, conducted the experiments, and wrote the manuscript; Kornienko MA, Letarov AV, Shitikov EA — planned the study, analyzed its results, and wrote the manuscript; Kuptsov NS, Efimov AD, Bogdan VI — conducted the experiments; Ilina EN — planned the study and wrote the manuscript.
Compliance with ethical standards: the experiments were conducted in full compliance with Biosafety Guidelines for working with risk group III–IV pathogens (SP 1.3.2322-08), Amendment 1 to Biosafety Guidelines for working with risk group III–IV pathogens (SP 1.3.2518-09), medical waste regulations (SanPin 2.1.7.279010), and Federal Clinical Guidelines on the rational use of bacteriophages for therapy and prevention of diseases.
Received: 2021-07-20
Accepted: 2021-08-25
Published online: 2021-09-22
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Fig. 1. Endotoxin concentrations in the purified vB_KpnM_Seu621 and vB_KpnP_Dlv622 phage lysates. 1 — crude lysate; 2 — precipitation with PEG; 3 — CsCl density gradient ultracentrifugation; 4 — sucrose density gradient ultracentrifugation; 5 — 100 kDa filtration; 6 — filtration through 0.22 µm filters in the presence of MgSO4
Fig. 2. Concentration of phage particles in purified vB_KpnM_Seu621 and vB_KpnP_Dlv622 lysates after purification. 1 — crude lysates; 2 — precipitation with PEG; 3 — CsCl density gradient ultracentrifugation; 4 — sucrose density gradient ultracentrifugation; 5 — 100 kDa filtration; 6 — filtration through 0.22 µm filters in the presence of MgSO4
Table. Number of doses of therapeutic vB_KpnM_Seu621 and vB_KpnP_Dlv622 phage cocktails and endotoxin content per dose calculated for 100 ml of the initial phage lysate
