NDLI: Evaluation of effectiveness of bacteriophage purification methods

Content Provider	Springer Nature : BioMed Central
Author	Binte Mohamed Yakob Adil, Siti Saleha Tucci, Joseph Irving, Helen Cianciarulo, Cassandra Kabwe, Mwila
Abstract	The use of bacteriophages for therapy has increased over the last decade. While there is need for clear regulatory pathways for bacteriophage approval for mainstream use in clinical practice, practitioners and patients have been able to access bacteriophage therapy under compassionate grounds and through magistral preparations. However, there is currently no standard for purifying these bacteriophages to ensure safety, and good manufacturing practice certification may not be achieved in these emergency uses. In this study, we employed an Interleukin Receptor Associated Kinase (IRAK) 3 knockout monocyte-based assay to evaluate the endotoxin removal efficacy of three common bacteriophage purification methods: Triton X-100 exposure, CsCl density gradient ultracentrifugation, and Pierce™ High-Capacity Endotoxin Removal Resin spin columns. In our experiments we tested these purification methods on three different bacteriophage morphotypes: siphovirus, podovirus and myovirus. We showed that the lowest endotoxin levels and immune responses were achieved when purifying bacteriophages with Triton-X treatment. The results from purifying with CsCl density gradient ultracentrifugation were comparable, and these were both significantly better than purification with Pierce™ High-Capacity Endotoxin Removal Resin spin columns. We also showed that Triton X-100 purification resulted in the lowest loss of bacteriophage titres. Finally, of the bacteriophages tested here, it did not appear that virus morphology affected efficacy of endotoxin removal.
Related Links	https://virologyj.biomedcentral.com/counter/pdf/10.1186/s12985-024-02580-y.pdf
Ending Page	10
Page Count	10
Starting Page	1
File Format	HTM / HTML
DOI	10.1186/s12985-024-02580-y
Journal	Virology Journal
Issue Number	1
Volume Number	21
Language	English
Publisher	BioMed Central
Publisher Date	2024-12-19
Access Restriction	Open
Subject Keyword	Virology Bacteriophage Endotoxin purification Triton X-100 Caesium chloride Endotoxin removal resin
Content Type	Text
Resource Type	Article
Subject	Virology Infectious Diseases
Journal Impact Factor	4/2023
5-Year Journal Impact Factor	3.8/2023

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Rapid isolation and purification of lactococcal bacteriophage DNA without the use of caesium chloride gradients

Synthesis of "Nereid," a new phenol-free detergent to replace Triton X-100 in virus inactivation.

Reconstitution of membranes with fractions of triton x- 100 which are easily removed. Anal biochem (1981).

The disruption of infectious bronchitis virus (IBV-41 strain) with triton X-100 detergent.

Solvent Extraction of Klebsiella pneumoniae Bacteriophage Lysates with 1-Dodecanol Results in Endotoxin Reduction with Low Risk of Solvent Contamination.

Production of highly immunogenic and safe Triton X-100 produced bacterial ghost vaccine against Shigella flexneri 2b serotype

Removal of endotoxin from protein solutions by phase separation using Triton X-114.

Sorption Equilibrium and Thermodynamics of Triton X-100 removal from aqueous Solutions

Removal of malachite green from water samples by cloud point extraction using Triton X-100 as non-ionic surfactant

Evaluation of effectiveness of bacteriophage purification methods

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Rapid isolation and purification of lactococcal bacteriophage DNA without the use of caesium chloride gradients

Synthesis of "Nereid," a new phenol-free detergent to replace Triton X-100 in virus inactivation.

Reconstitution of membranes with fractions of triton x- 100 which are easily removed. Anal biochem (1981).

The disruption of infectious bronchitis virus (IBV-41 strain) with triton X-100 detergent.

Solvent Extraction of Klebsiella pneumoniae Bacteriophage Lysates with 1-Dodecanol Results in Endotoxin Reduction with Low Risk of Solvent Contamination.

Production of highly immunogenic and safe Triton X-100 produced bacterial ghost vaccine against Shigella flexneri 2b serotype

Removal of endotoxin from protein solutions by phase separation using Triton X-114.

Sorption Equilibrium and Thermodynamics of Triton X-100 removal from aqueous Solutions

Removal of malachite green from water samples by cloud point extraction using Triton X-100 as non-ionic surfactant

Evaluation of effectiveness of bacteriophage purification methods