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Title:Phenotyping of mammalian CHO cell line in fed-batch bioprocess by intensified and classical procedure : master's thesis
Authors:ID Košir, Matic (Author)
ID Potočnik, Uroš (Mentor) More about this mentor... New window
ID Gaber, Rok (Co-mentor)
Files:.pdf MAG_Kosir_Matic_2023.pdf (8,88 MB)
MD5: 3B77D5E1C81B86006C8795A8565A5533
 
Language:English
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:Nowadays, pharmaceutical industry is striving to find more biologically relevant solutions and to move from synthetic to biological products. Biopharmaceuticals represent an emerging sector in the pharmaceutical industry and are useful for a wide range of indications, including oncology and rheumatology. The development and optimisation of bioprocesses at laboratory scale is crucial for process understanding and translation into production. Bioprocess development often begins with comparison of several potential clones to be characterized, tested in low-volume cultures with different media or other variables, and then evaluated for growth, productivity, and consistency. The purpose of this master's thesis is to review changes in growth curves, substrate consumption, yield, and product quality of a mammalian cell line when substituting glucose as the main carbon source with alternative energy sources. This master thesis presents an attempt to adapt a mammalian CHO cell line to alternative energy sources. A comparison of the cultivation with maintenance of the CHO mammalian cell line by an intensified and a classical process is also shown. Substrate requirements at different time points of the process were monitored using phenotypic microarrays. Based on the obtained and analyzed results of the phenotypic microarrays, we tried to optimize the cell culture supplementation strategy. We have tried to use small molecules to facilitate adaptation to alternative energy sources. Our results showed that growing CHO cell culture on an alternative energy source-maltose and cell reprogramming with small molecules affects the quality of the product. Trends in cell growth, substrate consumption, by-product excretion and final product concentration are similar between the reference cell cultures and those pre-treated with small molecules. We found that adapting a mammalian CHO cell line to an alternative energy source is a challenging process that would need to be carried out over a long period of time.
Keywords:CHO cells, bioprocess, phenotyping, alternative energy sources, glycan mapping
Place of publishing:Maribor
Place of performance:Maribor
Publisher:[M. Košir]
Year of publishing:2023
Number of pages:1 spletni vir (1 datoteka PDF (XI, 52 f.))
PID:20.500.12556/DKUM-85049 New window
UDC:604.4:615.32(043.2)
COBISS.SI-ID:171485955 New window
Publication date in DKUM:05.10.2023
Views:180
Downloads:73
Metadata:XML RDF-CHPDL DC-XML DC-RDF
Categories:KTFMB - FKKT
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Licences

License:CC BY 4.0, Creative Commons Attribution 4.0 International
Link:http://creativecommons.org/licenses/by/4.0/
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:19.09.2023

Secondary language

Language:Slovenian
Title:Fenotipizacija sesalske celične linije CHO v bioprocesu z dohranjevanjem po intenziviranem in klasičnem postopku
Abstract:Dandanes si farmacevtska industrija prizadeva najti bolj biološko ustrezne rešitve in preiti od sintetičnih k biološkim izdelkom. Biofarmacevtski izdelki so nastajajoči sektor v farmacevtski industriji in so uporabni za številne indikacije, vključno z onkologijo in revmatologijo. Razvoj in optimizacija bioprocesov v laboratorijskem merilu sta ključna za razumevanje procesov in prenos v proizvodnjo. Razvoj bioprocesov se pogosto začne s primerjavo več potencialnih klonov, ki jih je potrebno okarakterizirati, preizkusiti v kulturah majhnega obsega z različnimi mediji ali drugimi spremenljivkami ter nato oceniti rast, produktivnost in doslednost. Namen tega magistrskega dela je pregledati spremembe v krivuljah rasti, porabi substratov, produktivnosti in kakovosti izdelkov celične linije sesalcev pri zamenjavi glukoze kot glavnega vira ogljika z alternativnimi viri energije. To magistrsko delo predstavlja poskus prilagoditve celične linije CHO sesalcev alternativnim virom energije. Prikazana je tudi primerjava gojenja in vzdrževanja celične linije sesalcev CHO po intenziviranim in klasičnim postopkom. Potrebe po substratih v različnih časovnih točkah procesa smo spremljali z uporabo fenotipskih mikromrež. Na podlagi pridobljenih in analiziranih rezultatov fenotipskih mikromrež smo poskušali optimizirati strategijo dopolnjevanja celične kulture. Z uporabo malih molekul smo poskušali pohitriti adaptiranje na alternativne vire energije. Naši rezultati so pokazali, da gojenje celične kulture CHO na alternativnem viru energije - maltozi in reprogramiranje celic z majhnimi molekulami vpliva na kakovost produkta. Trendi rasti celic, porabe substratov, formacije stranskih produktov in koncentracije končnega izdelka so podobni med referenčnimi celičnimi kulturami in tistimi, ki so bile predhodno tretirane z majhnimi molekulami. Ugotovili smo, da je prilagajanje celične linije CHO sesalcev na alternativni vir energije zahteven proces, ki bi ga bilo treba izvajati v daljšem časovnem obdobju.
Keywords:CHO celice, bioproces, fenotipizacija, alternativni viri energije, mapiranje glikanov


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