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Phenotyping of mammalian CHO cell line in fed-batch bioprocess by intensified and classical procedure : master's thesis
Matic Košir, 2023, master's thesis

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
Published in DKUM: 05.10.2023; Views: 356; Downloads: 88
Full text (8,88 MB)

Primerjava bioprocesov s celično linijo CHO v različnih bioreaktorjih : diplomsko delo univerzitetnega študijskega programa I. stopnje
Matic Košir, 2020, undergraduate thesis

Abstract: Farmacevtska industrija dandanes stremi k iskanju biološko ustreznejših rešitev in prehodu iz sintetičnih k biološkim produktom. Biofarmacevtiki predstavljajo vzpenjajoči sektor v farmacevtski industriji in so uporabni za širok spekter indikacij, vključujoč onkologijo in revmatologijo. Razvoj in optimizacija bioprocesov na laboratorijski skali je ključnega pomena za poznavanje procesa in prenos v proizvodnjo. Centralni faktor za razvoj in proizvodnjo biološkega zdravila je bioreaktor. S svojo inovativno zasnovo in visokokakovostnimi materiali mora zagotavljati optimalno in stabilno okolje za gojenje in rast celic. Zadnja leta so v porastu bioreaktorji za enkratno uporabo. Različne študije so pokazale njihove prednosti, ki so pripomogle k nadaljnjim razvojem bioloških zdravil. Tako je prišlo do razvoja nove množice bioreaktorjev, ki se razlikujejo predvsem po obliki, vnosu moči, instrumentaciji in volumnu posode za kultivacijo. Namen raziskovalnega dela je bil izvesti 14-dnevni bioproces v 4-litrskem bioreaktorskem sistemu BIOne podjetja Distek in ga primerjati z referenčnima bioprocesoma. Prvi referenčni bioproces je bil izpeljan v mikrobioreaktorskem sistemu ambr® 250, drugi pa v 1000-litrskem bioreaktorju HyClone proizvajalca Thermo Scientific. Vse bioprocese smo izvedli z istim klonom celic in pod enakimi pogoji. Bioprocesne parametre smo spremljali s pomočjo različnih sond in analitskih naprav. Pridobljene meritve smo obdelali z različnimi statističnimi metodami in jih predstavili na diagramih. Osredotočili smo se na kinetiko rasti, metabolizem in produktivnost sesalskih celic. Rezultati kažejo, da je bioreaktorski sistem pri optimalnih pogojih zagotovil uspešno izvedbo bioprocesa s sesalsko celično linijo CHO. Ugotovili smo zadovoljivo primerljivost med bioprocesi. Na podlagi preučevanih koncentracij metabolitov in substratov, osmolalnosti, kinetiko rasti in koncentracijo produkta smo zaključili, da je omenjen bioreaktorski sistem primeren za nadaljnje študije na naši laboratorijski skali. V prihodnosti nameravamo stremeti k eksperimentom z drugimi celičnimi linijami in testiranju različnih parametrov. Uspešna izvedba bi nam prinesla zaupanje v ta bioreaktorski sistem in omogočila prevzem za širšo uporabo.
Keywords: bioreaktorji za enkratno uporabo, monoklonska protitelesa, biofarmacevtika, šaržni bioproces z dohranjevanjem, kinetika in metabolizem CHO celic.
Published in DKUM: 08.10.2020; Views: 1303; Downloads: 136
Full text (2,78 MB)

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