Phenotyping of mammalian CHO cell line in fed-batch bioprocess by intensified and classical procedure

master's thesis

Matic Košir (Author), Uroš Potočnik (Mentor), Rok Gaber (Co-mentor)

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;

Data

Language:	English
Year of publishing:	2023
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[M. Košir]
UDC:	604.4:615.32(043.2)
COBISS:	171485955
Views:	83
Downloads:	64
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	Slovenian
Secondary title:	Fenotipizacija sesalske celične linije CHO v bioprocesu z dohranjevanjem po intenziviranem in klasičnem postopku
Secondary 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.
Secondary keywords:	CHO celice;bioproces;fenotipizacija;alternativni viri energije;mapiranje glikanov;magistrske naloge;
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	1 spletni vir (1 datoteka PDF (XI, 52 f.))
ID:	19948642