Due to their narrow host range, bacteriophages offer a possible alternative to conventional antibiotic treatment of bacterial infections in humans. Phage-based control of pathogens is now already being used in agro-food sector. Apart from infection treatment and pathogen control, new bacteriophage applications such as drug and gene delivery system are being developed. Due to wide range of applications, the demand for bacteriophage production is expected to increase in the future. Downstream processes in phage production usually employ methods with low efficiency and poor scalability. The development of monolithic chromatografic media enabled the introduction of chromatography in the phage downstream processing and analytics. polyHIPE are a new class of monolithic chromatographic media that have not yet been tested for phage separation. The purpose of the master's thesis was to test the usability of differently functionalised polyHIPE chromatographic columns for phage and phage DNA separation and to optimise the chromatographic method. Using QA functionalised polyHIPE chromatographic column, we were able to separate T4 and T7 phage and their genomic DNA from phage lisate. The backpressure did not increase significantly even after a few consecutive cycles of binding and elution. Using optimised chromatographic method we analyzed the composition of phage lysate and monitor it's changing during the phage cultivation. The chromatographic method proved to be suitable for in-process control of phage cultivation. We found that pretreatment of phage lysate with PEG precipitation or the increase of Na+ concentration in loading sample can improve the resolution of the chromatographic method.
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