A non-chromatographic method for the removal of endotoxins from bacteriophages

Biotechnology and Bioengineering
Steven D BranstonEli Keshavarz-Moore

Abstract

The Ff filamentous bacteriophages show potential as a new class of therapeutics, displaying utility in materials science as well as pharmaceutical applications. These phages are produced by the infection of E. coli, a Gram-negative bacterium which unavoidably sheds endotoxins into the extracellular space during growth. Since endotoxin molecules are highly immunoreactive, separation from the phage product is of critical importance, particularly those developed for human therapeutic use. The properties of M13, one of the Ff group, present a purification challenge chiefly because the standard scalable method for endotoxin removal from proteins-anion exchange chromatography-is not applicable due to pI similarity between the particles. This article examines the potential of polyethylene glycol (PEG)-NaCl precipitation as a scalable method for the separation of endotoxins from phage M13. Precipitation of M13 by 2% (w/v) PEG 6 000, 500 mM NaCl reduced endotoxin contamination of the phage product by 88%, but additional precipitation rounds did not maintain this proportional decrease. Dynamic light scattering was subsequently used to determine the effectiveness of a detergent to disassociate endotoxin molecules from M13. As a result, PE...Continue Reading

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Citations

Jun 16, 2015·Frontiers in Microbiology·Eyal Dor-On, Beka Solomon
Jun 22, 2019·Scientific Reports·Sidharth RazdanSutapa Barua
Apr 26, 2020·Biotechnology and Bioengineering·Mason SchneierSutapa Barua
Nov 28, 2016·Applied Microbiology and Biotechnology·Douglas B FigueiredoViviane Maimoni Gonçalves
Nov 2, 2019·Microbiology and Molecular Biology Reviews : MMBR·Krystyna Dąbrowska, Stephen T Abedon
Apr 2, 2020·Nature Nanotechnology·Daniel LausterChristian P R Hackenberger
Oct 20, 2020·Frontiers in Microbiology·Yoon Jung Hwang, Heejoon Myung
Apr 26, 2021·Biotechnology Advances·Jorge JoãoAna M Azevedo
Aug 26, 2017·Microbiology Spectrum·Stephen T Abedon

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