20.500.12556/RUL-160299 Ločevanje bakteriofagov na kromatografskih nosilcih polyHIPE s funkcionalnima skupinama QA in DEAE Separation of bacteriophages on chromatographic polyHIPE supports with functional groups QA and DEAE Kromatografska izolacija bakteriofagov, predvsem za uporabo pri fagni terapiji, predstavlja izjemno pomemben vidik s strani varnosti in učinkovitosti njihove uporabe. Glavni namen diplomskega dela je bil pokazati možnost ločevanja bakteriofagov, po principu ionsko izmenjevalne kromatografije, z uporabo polyHIPE kromatografskih nosilcev. Bakteriofage smo nagojili, izvedli korak predhodnega čiščenja s PEG in jih od bakterijskega lizata kromatografsko ločili na sistemu HPLC. Dokazali smo, da je ločevanje bakteriofagov T7 in ɸ6, z uporabo kromatografskih nosilcev polyHIPE, mogoče. Močan anionski izmenjevalec QA se je v primerjavi s šibkim anionskim izmenjevalcem DEAE, izkazal za učinkovitejšega pri ločevanju obeh vrst bakterofagov, predvsem zaradi večjih izkoristkov oziroma manjših izgub med procesom. Primerjava med bakteriofagoma je pokazala, da je kromatografija faga T7, tudi v tem primeru zaradi večjega izkoristka učinkovitejša od izolacije virusa ɸ6. Za slednjega je značilen tudi neučinkovit postopek obarjanja s PEG, kakor tudi večja težavnost dela med celotnim eksperimentom. Chromatographic isolation of bacteriophages, especially for the application of phage therapy, represents an extremely important aspect in terms of the safety and efficacy of their use. The main aim of this thesis was to demonstrate the possibility of separating bacteriophages based on a principle of ion-exchange chromatography, using polyHIPE chromatographic supports. We cultivated bacteriophages, performed a preliminary purification step with PEG, and chromatographically separated them from the bacterial lysate using an HPLC system. We demonstrated that the separation of bacteriophages T7 and ɸ6 using polyHIPE chromatographic supports is feasible. The strong anion exchanger QA proved to be more effective in comparison to the weak anion exchanger DEAE for the separation of both types of bacteriophages, primarily due to higher yields and lower losses during the process. Comparing two bacteriophages, the chromatography of phage T7 was found to be more efficient than isolation of phage ɸ6, again due to higher yield. For the latter, an ineffective PEG precipitation process and more difficult handling throughout the isolation process was noticed. bakteriofagi ionsko izmenjevalna kromatografija polyHIPE bacteriophages ion-exchange chromatography polyHIPE true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2024-08-25 07:16:10 2024-08-25 07:16:14 2024-10-03 13:47:01 0000-00-00 00:00:00 2024 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 241760 205490435 40705.pdf 40705.pdf 1 4C85B04139EE2127B6888929299EB6B4 33ea9ea2b13182c950a34990104874e640ed35c1cf945c139a5db422606d9ebe edd4fca0-62a0-11ef-8f74-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=188879 Biotehniška fakulteta 0 0 0