magistrsko delo
Manca Volk (Author), Marjanca Starčič Erjavec (Reviewer), Uroš Petrovič (Mentor), Uroš Petrovič (Thesis defence commission member), Matjaž Peterka (Thesis defence commission member), Marjanca Starčič Erjavec (Thesis defence commission member), Rok Kostanjšek (Thesis defence commission member), Matjaž Peterka (Co-mentor)

Abstract

Pridobivanje plazmidnih vektorjev za uporabo v genski terapiji je odvisno od donosnosti pridobivanja, ki mora zagotoviti zadostne količine plazmidne DNA v obliki kovalentno zaprtega kroga. Na količino in kakovost plazmidne DNA v zaključnih procesih vpliva več dejavnikov, ki jih je potrebno optimizirati v začetnih, pripravljalnih fazah bioprocesa. Z namenom povišanja donosnosti bioprocesa smo v sklopu magistrske naloge optimizirali pripravljalni proces. Z gojenjem smo pričeli v 3-litrskem bioreaktorskem sistemu, kjer smo lahko tekom gojenja spremljali rast, parcialni tlak kisika ter pH v kontroliranem okolju. Gojenje smo najprej izvedli v šaržu brez dohranjevanja in kasneje v šaržu z dohranjevanjem. Testirali smo vpliv parcialnega tlaka kisika ter temperature na količino plazmidne DNA. Z uporabo šaržnega gojenja z dohranjevanjem smo dosegli 79,5 % višji donos plazmidne DNA v primerjavi s šaržnim gojenjem brez dohranjevanja.

Keywords

plazmidna DNA;genska terapija;pridobivanje plazmidne DNA;optimizacija bioprocesa;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [M. Volk]
UDC: 577.2(043.2)
COBISS: 19055363 Link will open in a new window
Views: 632
Downloads: 232
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Other data

Secondary language: English
Secondary title: Optimization of production and isolation of plazmid DNA for gene therapy
Secondary abstract: The manufacturing of plasmid vectors for gene therapy applications depends on the yield of plasmid DNA production, which has to be high enough to provide sufficient amount of super coiled plasmid DNA. In the downstream process the amount and quality of plasmid DNA are affected by many factors, which have to be optimised in the upstream process of bioprocess development. The scope of the thesis was to increase the yield of the upstream process. Bacterial cultivation began in a 3 L bioreactor vessel where the growth of bacteria, pH and oxygen partial pressure were monitored in a controlled environment. Bacterial cultivation was carried out a batch mode and also in a fed-batch mode. The effect of oxygen partial pressure and temperature on the production of plasmid DNA was tested. The use of fed-batch mode increased the yield of plasmid DNA by 79.5% compared to batch mode.
Secondary keywords: plasmid DNA;gene therapy;plasmid DNA production;bioprocess optimisation;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. Ljubljana, Biotehniška fak.
Pages: X, 37, [1] f.
ID: 11688439