Bacteriophages are viruses that infect bacteria. Nowadays bacteriophage therapy is gaining interest, as more and more antibiotic-resistance bacteria appears. The aim of the master’s thesis was to test different formulation strategies. We used two bacteriophages: a bacteriophage for Klebsiella pneumoniae (KP49) and a bacteriophage for Escherichia coli (S18036). To test the formulation, we first tested the propagation of bacteriophages on a smaller scale, and then scale up the process for propagation in the bioreactor. After multiplication, the bacteriophages were concentrated and diafiltered by tangential flow filtration (TFF) through a membrane with a pore size of 100 kDa. Different bacteriophage formulations were prepared, in solution and powder. For liquid formuation, we tested the effect of adding the excipient or magnesium ion, on the stability of the bacteriophages. We also tested the effect of storage temperature on the concentration of functional bacteriophages and observe the difference between bacteriphage KP49 and S18036, with the latter being more stable. For powder fomulation, formulation with spray drying was used, where we first tested the effect of the temperature on yield. The best parameter was selected, in which we tested storage at different temperatures at different timepoints. The conclusion was simillar to the liquid fomulations, with bacteriophage S18036 being more resistant in comparison with KP49. We concluded that bacteriphage KP49 had better survival in liquid formulations, while bacteriphage S18036 had comparable survival in all formulations tested. Based on the results, we concluded that a certain formulation is not generally more suitable for all bacteriophages, but it is necessary to determine the most suitable one for a specific bacteriophage.
|
