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Ločevanje bakteriofagov na kromatografskih nosilcih polyHIPE s funkcionalnima skupinama QA in DEAE
ID Jereb, Nik (Author), ID Podgornik, Aleš (Mentor) More about this mentor... This link opens in a new window

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Abstract
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.

Language:Slovenian
Keywords:bakteriofagi, ionsko izmenjevalna kromatografija, polyHIPE
Work type:Bachelor thesis/paper
Organization:BF - Biotechnical Faculty
Year:2024
PID:20.500.12556/RUL-160299 This link opens in a new window
COBISS.SI-ID:205490435 This link opens in a new window
Publication date in RUL:25.08.2024
Views:80
Downloads:12
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Secondary language

Language:English
Title:Separation of bacteriophages on chromatographic polyHIPE supports with functional groups QA and DEAE
Abstract:
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.

Keywords:bacteriophages, ion-exchange chromatography, polyHIPE

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