Ababi, Maria (2022) Engineering bacteriophage P2 to expand its antimicrobial activity. PhD thesis, University of Warwick.

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Abstract

Phage therapy is a promising alternative to antibiotics for treating multi-drug resistant bacterial infections. Phage therapy is however currently only used on a compassionate basis. The main aim in this thesis was thus to develop a phage-based platform able to produce phage-like particles for practical therapeutic applications, taking into account the following key challenges: regulatory framework, safety, stability, mass production and flexibility for practical applications. For this, the broad-spectrum phage P2 was engineered, taking advantage of its relationship with satellite phage P4, to produce P4-like particles carrying ~4 kb custom cargo with therapeutic properties. After optimization, the production resulted in about 10^11 helper-free P4-like particles per mL. For informed downstream bioprocessing, the particles were characterised as: hydrophilic, weighing about 23.54 MDa, with isoelectric point between pH 4.2 and 4.7. For safe preclinical trials, the endotoxins were removed by affinity chromatography down to 0.25 endotoxin units per 10^9 particles and the particles stability was confirmed for one month at 2-8 °C. The cargo capacity was also extended to up to 25.8 kb by deleting the P4 sid gene. The flexibility of the engineering platform was then investigated by testing two proof of concepts (“prime only”, “prime and kill”) and by testing the feasibility for developing a directed evolution (DE) platform for pyocin-like particles.

Item Type:	Thesis (PhD)
Subjects:	Q Science > QR Microbiology R Medicine > RM Therapeutics. Pharmacology
Library of Congress Subject Headings (LCSH):	Bacteriophages -- Therapeutic use, Bacterial diseases -- Treatment, Anti-infective agents -- Development, Drug resistance in microorganisms
Official Date:	October 2022
Dates:	Date Event October 2022 UNSPECIFIED
Institution:	University of Warwick
Theses Department:	Warwick Medical School
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	Jaramillo, Alfonso (Jaramillo Rosales) ; Jameson, Eleanor
Sponsors:	Medical Research Council (Great Britain) ; Spain. Ministerio de Ciencia e Innovación ; GlaxoSmithKline
Format of File:	pdf
Extent:	371 pages
Language:	eng

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