magistrsko delo
Gašper Dokl (Author), David Stopar (Reviewer), Uroš Petrovič (Mentor), Franc Smrekar (Co-mentor)

Abstract

Bakteriofagi so virusi, ki gostujejo v bakterijah. Dandanes je zamisel o zdravljenju bakterijskih okužb z bakteriofagi znova postala aktualna, saj se pojavlja vse več proti antibiotikom odpornih bakterij. Cilj magistrske naloge je bil testiranje različnih strategij formulacij. Uporabili smo dva bakteriofaga: bakteriofag za bakterijo Klebsiella pneumoniae (KP49) in bakteriofag za bakterijo Escherichia coli (S18036). Za testiranje formulacije smo najprej testirali namnoževanje bakteriofagov v manjšem obsegu, rezultate pa nato uporabili za postopek namnoževanja v bioreaktorju. Po namnoževanju smo bakteriofage koncentrirali in diafiltrirali s tangencialno filtracijo (TFF; angl. tangential flow filtration) skozi membrano z velikostjo por 100 kDa. Nato smo pripravili različne bakteriofagne pripravke; v raztopini oziroma posušene pripravke. Pri pripravkih v raztopini smo testirali vpliv dodajanja kompatibilnih topljencev oziroma magnezijevega iona na stabilnost bakteriofagov. Ugotavljali smo tudi učinek temperature shranjevanja na koncentracijo funkcionalnih bakteriofagov in opazili razlike med bakteriofagoma KP49 in S18036, pri čemer je bil slednji bolj stabilen. Pri testiranju posušenega pripravka smo uporabili sušenje z razprševanjem, kjer smo najprej testirali vpliv temperature na izkoristek. Iz testiranih pogojev smo izbrali temperaturo sušenja z najboljšim izkoristkom, to je 60 °C. Nato smo s sušenjem pri 60 °C pridelali bakteriofagni pripravek v prahu in pri različnih časih testirali shranjevanje pri različnih temperaturah. Podobno kot pri formulacijah v raztopini smo ugotovili, da je bakteriofag S18036 odpornejši od KP49. Ugotovili smo, da je pri bakteriofagu KP49 preživetje boljše pri pripravkih v raztopini, medtem ko je preživetje bakteriofaga S18036 primerljivo pri vseh formulacijah. Na podlagi rezultatov ugotavljamo, da določena formulacija ni splošno primernejša za vse bakteriofage, ampak je potrebno najprimernejšo določiti za specifičen bakteriofag.

Keywords

bakteriofagi;formulacija;sušenje z razprševanjem;formulacija v raztopini;fagna terapija;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [G. Dokl]
UDC: 602.3:578.347
COBISS: 219138307 Link will open in a new window
Views: 26
Downloads: 441
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Other data

Secondary language: English
Secondary title: ǂThe ǂinfluence of different bacteriophage formulation strategies on their stability
Secondary abstract: 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.
Secondary keywords: bacteriophages;formulation;spray drying;liquid formulation;phage therapy;
Type (COBISS): Master's thesis/paper
Study programme: 0
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za mikrobiologijo
Pages: 1 spletni vir (1 datoteka PDF (XII, 47 str., [3] f. pril.))
ID: 25518732