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Open AccessArticle

Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol

1
Department of Immunology and Molecular Microbiology, Texas Tech University Health Sciences Center, 3601 4th Street, Lubbock, TX 79430, USA
2
Roche Molecular Systems, 983 University Avenue B200, Los Gatos, CA 95032, USA
3
Center for Infectious Disease, Department of Molecular Biosciences, The University of Texas at Austin, 1 University Station A5000, Austin, TX 78712, USA
*
Authors to whom correspondence should be addressed.
Viruses 2019, 11(3), 241; https://doi.org/10.3390/v11030241
Received: 21 January 2019 / Revised: 27 February 2019 / Accepted: 8 March 2019 / Published: 11 March 2019
(This article belongs to the Special Issue Biotechnological Applications of Phage and Phage-Derived Proteins)
The ‘Appelmans protocol’ is used by Eastern European researchers to generate therapeutic phages with novel lytic host ranges. Phage cocktails are iteratively grown on a suite of mostly refractory bacterial isolates until the evolved cocktail can lyse the phage-resistant strains. To study this process, we developed a modified protocol using a cocktail of three Pseudomonas phages and a suite of eight phage-resistant (including a common laboratory strain) and two phage-sensitive Pseudomona aeruginosa strains. After 30 rounds of selection, phages were isolated from the evolved cocktail with greatly increased host range. Control experiments with individual phages showed little host-range expansion, and genomic analysis of one of the broad-host-range output phages showed its recombinatorial origin, suggesting that the protocol works predominantly via recombination between phages. The Appelmans protocol may be useful for evolving therapeutic phage cocktails as required from well-defined precursor phages. View Full-Text
Keywords: Appelmans; bacteriophage evolution; bacteriophage recombination; phage therapy; Pseudomonas aeruginosa; antibiotic resistance Appelmans; bacteriophage evolution; bacteriophage recombination; phage therapy; Pseudomonas aeruginosa; antibiotic resistance
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Burrowes, B.H.; Molineux, I.J.; Fralick, J.A. Directed in Vitro Evolution of Therapeutic Bacteriophages: The Appelmans Protocol. Viruses 2019, 11, 241.

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