20.500.12556/RUL-125513 Vpeljava procesne analitične tehnologije v bioprocese s sesalskimi celicami Implementation of process analytical technology in bioprocessing with mammalian cell cultures Biofarmacevtska industrija stremi k boljšemu nadzoru nad procesi v proizvodnji zdravil. Namen naloge je bil preveriti možnost vpeljave procesne analitične tehnologije (PAT) v laboratorijske bioprocese s sesalskimi celicami ter opredeliti uporabnost predstavljene tehnologije za boljši nadzor nad bioprocesi. Klasično ''off-line'' metodo spremljanja rasti celic z barvanjem in štetjem smo primerjali z ''on-line'' metodo spremljanja rasti preko kapacitivnosti s sondami ABER. Skupno smo izpeljali dvanajst bioprocesov v 5-litrskih bioreaktorjih za enkratno uporabo, kjer smo dvanajst dni gojili dve različni celični liniji sesalskih celic CHO. Spremljali smo kapacitivnost skozi celoten proces ter dnevno vzorčili celično kulturo na napravi Vi-Cell. Rezultati so pokazali na visoke stopnje korelacije med obema metodama spremljanja gostote živih celic, sploh v začetnih fazah rasti. Visoke korelacije med meritvami smo zabeležili tudi v časovnem območju temperaturnih premikov. Narejen je bil tudi regresijski model za napovedovanje temperaturnih in pH premikov na podlagi kapacitivnosti. Pokazali smo, da lahko pod primernimi pogoji s pomočjo modeliranja uspešno napovemo in avtomatiziramo premike za določeno celično linijo. Opredelili smo se tudi do vpliva različnih faktorjev na meritve kapacitivnosti, kot sta velikost in viabilnost celic. Z rezultati smo potrdili vse postavljene hipoteze o primernosti uporabe ABER sond za uspešno in natančno spremljanje rasti celic ''on-line''. Opravljeno delo prispeva k napredovanju vpeljave PAT tehnologije v bioprocese s sesalskimi celicami na laboratorijski ravni. Biopharmaceutical industry aims for better surveilance over processes in drug manufacturing. The purpose of this work was to assess the possibilities of implementing process analytical technology (PAT) in laboratory scale bioprocessing with mammalian cell cultures and to determine the value of the introduced technology in better surveilance over the bioprocesses. We compared the classic ''off-line'' methods of cell growth monitoring by dyeing and counting with the ''on-line'' cell growth monitoring via capacitance measuring with ABER probes. The experiment was conducted with twelve bioprocesses in 5 litre single-use bioreactors. Two different CHO cell lines were grown there for twelve days. We monitored the capacitance throughout the entire process. We sampled and analyzed the culture daily on Vi-Cell cell counter. Results have shown high degrees of correlation between two methods of cell growth monitoring, especially during the early stages. High correlation betwen measurments was also observed during the temperature shift. A regression model for predicting temperature and pH shifts via capacitance was also made. We have shown that we can successfully predict and automatize shifts for a specific cell line under the right conditions by modeling. We have also expressed our thoughts about the impact of various factors like cell size and viability on capacitance measurements. Our current results confirm all of our hypotheses of ABER probes usage for successful and accurate ''on-line'' monitoring of cell growth. The work done contributes in advancing the implementation of PAT in laboratory scale bioprocessing with mammalian cell cultures. procesna analitična tehnologija kapacitivnost bioproces sesalske celice korelacija process analytical technology capacitance bioprocess mammalian cell culture correlation true false false [S. Cundrič] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2021-03-21 07:15:29 2021-03-21 07:15:41 2022-09-05 03:51:07 0000-00-00 00:00:00 2018 0 Ljubljana 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 602.1:53.082.9:606:62(043.2) 191060 61828611 12499.pdf 12499.pdf 1 EC277BCBD0EEB0585DC4506710776C03 9b468b147eb92bb0fb558ac58ad32ed7d510c2ea2c0a293df2d8a487d2079eac b989dda0-a1bb-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=141253 Biotehniška fakulteta 0 0 0