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Vpeljava procesne analitične tehnologije v bioprocese s sesalskimi celicami
ID Kavčič, Urban (Author), ID Podgornik, Aleš (Mentor) More about this mentor... This link opens in a new window, ID Starcevic, Štefan (Co-mentor)

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Abstract
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.

Language:Slovenian
Keywords:procesna analitična tehnologija, kapacitivnost, bioproces, sesalske celice, korelacija
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[S. Cundrič]
Year:2018
PID:20.500.12556/RUL-125513 This link opens in a new window
UDC:602.1:53.082.9:606:62(043.2)
COBISS.SI-ID:61828611 This link opens in a new window
Publication date in RUL:21.03.2021
Views:1160
Downloads:102
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Secondary language

Language:English
Title:Implementation of process analytical technology in bioprocessing with mammalian cell cultures
Abstract:
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.

Keywords:process analytical technology, capacitance, bioprocess, mammalian cell culture, correlation

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