20.500.12556/RUL-112852 Ugotavljanje ustreznosti regeneracije in čiščenja kromatografskih kolon v procesu izolacije biološke učinkovine viEvaluation of chromatographic columns regeneration and cleaning efficiency during drug substance (biomolecule) isolation process Proizvodnja bioloških zdravil vključuje več korakov izolacije in čiščenja aktivne učinkovine. Za čiščenje bioloških učinkovin (biomolekul) so najpogosteje v rabi različne kromatografske tehnike, pri katerih kromatografske kolone predstavljajo osnovni del opreme. Po vsakem industrijskem procesu (seriji) sta ključnega pomena regeneracija in čiščenje kolone, saj s tem preprečimo prenos produkta iz ene serije v drugo. Namen diplomske naloge je bil preveriti učinkovitost regeneracije in čiščenja kromatografskih kolon v procesu čiščenja biološke učinkovine. V ta namen smo po rednem procesu regeneracije izvedli elucijo brez predhodnega nanosa produkta ter dobljeni kromatogram primerjali s tipičnim kromatogramom, značilnim za posamezno kromatografsko stopnjo. Poleg spremljanja absorbance med to stopnjo, smo v eluatu izmerili tudi koncentracijo biološke učinkovine. Rezultati so potrdili hipotezo, ki predvideva, da sta regeneracija in čiščenje kromatografskih kolon v procesu proizvodnjebiološke učinkovine učinkoviti. Test smo izvedli na vseh kromatografskih stopnjah v proizvodnji biološke učinkovine z izjemo gelske izključitvene kromatografije, saj pri tejkromatografiji ne prihaja do interakcij med produktom in kromatografskim nosilcem. Production of biological drugs involves multiple steps of isolation and purification of the target drug substance (biomolecule). Most commonly used technique for isolation and purification is chromatography of which chromatographic columns are the vital part. After each industrial process, regeneration and cleaning of chromatographic columns are crucial to prevent carry-over of the product from one batch to another. The aim of this diploma thesis was therefore to test the efficiency of regeneration and cleaning of chromatographic columns that are used in production of target biomolecule. For this purpose a blank elution (without previously loading the protein) was executed after a routine process. The chromatogram of blank elution was compared to the master chromatogram, which showed no signal of absorbance. In addition, the content of the drug substance in blank eluate was analysed. Results obtained in this study confirmed the hypothesis, that regeneration and cleaning of chromatographic columns involved in drug substance production are effective. The study was performed at all chromatographic steps in drug substance production with the exception of the size exclusion chromatography, where there is no interaction of product with chromatographic resin. biološke učinkovine zaključni procesi ionska kromatografija hidrofobna kromatografija procesna analitika drug substance final processing product carry-over between batches ion exchange chromatography hydrophobic interaction chromatography process analytics true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2019-11-15 13:55:06 2019-11-15 13:55:07 2022-08-25 03:51:38 0000-00-00 00:00:00 2019 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 5078 1538497219 Andelkovic_Marko_-_Ugotavljanje_ustreznosti_regeneracije_in_ciscenja_kromatografskih_kolon_v_pro.pdf Andelkovic_Marko_-_Ugotavljanje_ustreznosti_regeneracije_in_ciscenja_kromatografskih_kolon_v_pro.pdf 1 A8A29578F2AC56619D67DD986FD1CE90 2f61acb30884e5581cf7d82ef213780b07ac5b1f61c18012acf2900b36ac67ea f8132eb1-a1b7-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=124791 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0