The therapeutic use of lytic bacteriophages has a potential as a viable solution to the worldwide problem of increasing number of drug-resistant bacteria (1). In recent years, there has been a great emphasis towards the modification of bacteriophage particles for their use as a nanomaterial for diagnostic and therapeutic applications, vaccines or for systems employed in bacteria detection.

This study concerns physicochemical properties of bacteriophage T4, purified using the extraction method (2) and membrane filtration. The purification process allowed removal of endotoxins, culture medium components and residues generated after disintegration of bacterial cells. The titer of bacteriophage T4 in purified preparation was 4,2x10¹³ pfu/ml and endotoxin level, determined by Limulus Amebocyte Assay, was 76 EU/ml. The effect of ionic strength on stability of bacteriophage suspension was studied using atomic force microscopy (AFM) and dynamic light scattering (DLS) measurement. On the basis of size measurements it was found that critical level of sodium bicarbonate concentration, below which the process of size increase starts, is about 0.03 M. The AFM imaging indicates that increase in particle sizes can be linked to the process of aggregation of bacteriophage particles emerging in the alkaline environment of lower ionic strength.

Presented results prove that physicochemical properties of bacteriophage T4 depend on the medium composition, in particular on the ionic strength and pH.

This project was supported by the National Centre for Research and Development, Poland (Grant No 13-0089-06).

References:

1. Parisien A, Allain B, Zhang J, Mandeville R, Lan CQ (2008) Novel alternatives to antibiotics: bacteriophages, bacterial cell wall hydrolases, and antimicrobial peptides. J Appl Microbiol 104: 1-13
2. Boratyński J, Szermer-Olearnik B, Weber-Dąbrowska B, Górski A Sposób uzyskiwania preparatów bakteriofagowych obejmujący namnażanie bakteriofagów w komórkach bakterii, poddawanie lizie komórek z bakteriofagami. Polish Patent no 213388 PL

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